This document discusses immobilization of biomolecules on biomaterial surfaces. It begins by explaining that biomaterials must have suitable bulk and surface properties to function in biological environments. Common approaches involve fabricating materials with adequate bulk properties, then modifying the surface to enhance biocompatibility. The document then discusses various biomolecules that can be immobilized on surfaces like proteins, peptides, polysaccharides and describes techniques for covalent and non-covalent immobilization. Specific examples of immobilizing collagen, RGD peptide, hyaluronic acid and other biomolecules on scaffolds are provided to support tissue engineering applications.
Enzyme immobilization involves confining enzyme molecules to a distinct phase separate from the reaction substrates and products. This protects enzymes and allows for continuous use. Common immobilization methods include adsorption, ionic binding, covalent binding, cross-linking, and entrapment. Adsorption and ionic binding rely on weak interactions, while covalent binding forms stronger covalent bonds. Entrapment encloses enzymes within a semi-permeable polymer membrane or matrix. Immobilization can increase enzyme stability but may also reduce activity depending on the method used and how it affects enzyme structure and substrate binding/product release.
Protein purification techniques can be categorized into those based on molecular size, solubility, and electric charge. Size-based techniques include dialysis, ultrafiltration, and size-exclusion chromatography which separate proteins based on their ability to pass through semi-permeable membranes or porous beads. Solubility-based techniques include isoelectric precipitation and salting out which alter a protein's solubility by adjusting pH or salt concentration. Charge-based techniques such as ion-exchange and electrophoresis separate proteins using their net electric charge in an applied electric field or ion-exchange column.
This document discusses protein folding and aggregation. It describes the primary, secondary, tertiary, and quaternary structures of proteins. Protein folding is guided by amino acid sequence and cellular environment. Misfolding can occur and result in non-native protein conformations. This leads to protein aggregation and the formation of amyloid fibrils or inclusion bodies. Protein aggregation is associated with neurodegenerative diseases like Alzheimer's and Parkinson's. Factors like sequence, environment, concentration, pH, and temperature can affect protein aggregation. Understanding protein aggregation may help develop therapies for related diseases.
A biosensor is a device that uses biological components like enzymes, antibodies, or living cells to detect analytes. It consists of a biological recognition element and a physicochemical transducer. A nanobiosensor is a biosensor that operates on the nanoscale. Some key applications of nanobiosensors include detecting DNA, proteins, cells, and biomarkers for medical diagnostics. They can also be used for environmental monitoring, food safety testing, and other areas. Despite their potential, commercialization of biosensors has faced challenges related to biomolecule immobilization, device sensitivity and reproducibility, and cost-effectiveness.
Biosensors (structure and application)Tarun Kapoor
This document discusses biosensors, including their structure, history, working principles, types, sensing elements, transducers, amplifiers, and applications. It defines a biosensor as a device that combines a biological component with a physicochemical detector to detect analytes. The key components are a biological recognition element, transducer to convert the biological response into a measurable signal, and amplifier. Major applications mentioned include glucose monitoring, environmental monitoring, drug discovery, and food/agriculture testing. Disadvantages include inability to use heat sterilization and stability issues with biological materials.
A biosensor is a device that uses biological components like enzymes or antibodies to detect analytes. It converts a biological response into an electrical signal. Some key points:
- Glucose biosensors are widely used by diabetics to monitor blood glucose levels. They detect glucose using the enzyme glucose oxidase.
- Biosensors have applications in food freshness monitoring, environmental monitoring, medical diagnostics, agriculture, and more.
- Common types include electrochemical, optical, thermal, and piezoelectric biosensors. Electrochemical biosensors for glucose are a commercial success story.
This document summarizes the application of nanotechnology to regenerative medicine. It begins with background on regenerative medicine and its focus on repairing damaged tissues through stem cell therapy and tissue engineering. It then discusses how nanotechnology can help because biological processes occur on the nanoscale. Examples are given of FDA-approved regenerative products showing the commercial potential. Developments in regenerative medicine using stem cells and gene therapy for various diseases are also outlined. In general, the document shows how nanotechnology may improve regenerative approaches by better interacting with cellular structures and components that function on the nanoscale level.
Enzyme immobilization involves confining enzyme molecules to a distinct phase separate from the reaction substrates and products. This protects enzymes and allows for continuous use. Common immobilization methods include adsorption, ionic binding, covalent binding, cross-linking, and entrapment. Adsorption and ionic binding rely on weak interactions, while covalent binding forms stronger covalent bonds. Entrapment encloses enzymes within a semi-permeable polymer membrane or matrix. Immobilization can increase enzyme stability but may also reduce activity depending on the method used and how it affects enzyme structure and substrate binding/product release.
Protein purification techniques can be categorized into those based on molecular size, solubility, and electric charge. Size-based techniques include dialysis, ultrafiltration, and size-exclusion chromatography which separate proteins based on their ability to pass through semi-permeable membranes or porous beads. Solubility-based techniques include isoelectric precipitation and salting out which alter a protein's solubility by adjusting pH or salt concentration. Charge-based techniques such as ion-exchange and electrophoresis separate proteins using their net electric charge in an applied electric field or ion-exchange column.
This document discusses protein folding and aggregation. It describes the primary, secondary, tertiary, and quaternary structures of proteins. Protein folding is guided by amino acid sequence and cellular environment. Misfolding can occur and result in non-native protein conformations. This leads to protein aggregation and the formation of amyloid fibrils or inclusion bodies. Protein aggregation is associated with neurodegenerative diseases like Alzheimer's and Parkinson's. Factors like sequence, environment, concentration, pH, and temperature can affect protein aggregation. Understanding protein aggregation may help develop therapies for related diseases.
A biosensor is a device that uses biological components like enzymes, antibodies, or living cells to detect analytes. It consists of a biological recognition element and a physicochemical transducer. A nanobiosensor is a biosensor that operates on the nanoscale. Some key applications of nanobiosensors include detecting DNA, proteins, cells, and biomarkers for medical diagnostics. They can also be used for environmental monitoring, food safety testing, and other areas. Despite their potential, commercialization of biosensors has faced challenges related to biomolecule immobilization, device sensitivity and reproducibility, and cost-effectiveness.
Biosensors (structure and application)Tarun Kapoor
This document discusses biosensors, including their structure, history, working principles, types, sensing elements, transducers, amplifiers, and applications. It defines a biosensor as a device that combines a biological component with a physicochemical detector to detect analytes. The key components are a biological recognition element, transducer to convert the biological response into a measurable signal, and amplifier. Major applications mentioned include glucose monitoring, environmental monitoring, drug discovery, and food/agriculture testing. Disadvantages include inability to use heat sterilization and stability issues with biological materials.
A biosensor is a device that uses biological components like enzymes or antibodies to detect analytes. It converts a biological response into an electrical signal. Some key points:
- Glucose biosensors are widely used by diabetics to monitor blood glucose levels. They detect glucose using the enzyme glucose oxidase.
- Biosensors have applications in food freshness monitoring, environmental monitoring, medical diagnostics, agriculture, and more.
- Common types include electrochemical, optical, thermal, and piezoelectric biosensors. Electrochemical biosensors for glucose are a commercial success story.
This document summarizes the application of nanotechnology to regenerative medicine. It begins with background on regenerative medicine and its focus on repairing damaged tissues through stem cell therapy and tissue engineering. It then discusses how nanotechnology can help because biological processes occur on the nanoscale. Examples are given of FDA-approved regenerative products showing the commercial potential. Developments in regenerative medicine using stem cells and gene therapy for various diseases are also outlined. In general, the document shows how nanotechnology may improve regenerative approaches by better interacting with cellular structures and components that function on the nanoscale level.
Application of Biological Assemblies in NanoBiotechnology PPtZohaib HUSSAIN
The document discusses several applications of biological assemblies in nanobiotechnology. It describes how biological assemblies, such as hemoglobin, are the functional forms of molecules and can be used to build larger structures. It then discusses using drug nanocrystals to improve drug solubility and bioavailability. Next, it covers using nano-containers like liposomes for targeted drug delivery. The document also discusses using protein crystals called S-layers for nanolithography and peptide templates for controlling biomineralization. Finally, it discusses the potential of utilizing biomineralization principles in nanotechnology applications like tissue engineering.
Immobilised enzymes ppt presentation - dr. r. mallikamallikaswathi
The document discusses immobilized enzymes and their applications. It defines immobilization as imprisoning cells or enzymes onto a support matrix. The first immobilized enzyme was amino acylase used in Japan. Immobilization offers benefits like enhanced efficiency, reproducibility, and stability. Common supports discussed include natural polymers like alginate and chitin, and inorganic materials like zeolites. The main immobilization methods covered are adsorption, covalent bonding, entrapment, and encapsulation. Important industrial uses of immobilized enzymes include production of antibiotics, beverages, amino acids, and their use in biomedical applications like disease diagnosis.
Biosensors are the analytical device that are used to measure the concentration of analye , these type of biosensors are made with conjugation of enzymes as a biological eliment to quantify a (bio)chemical substance / analyte are reffered to as Enzyme-probe Biosensors .
Biosensors are of many types but focusing on Enzyme biosensors there are 4 main types which are briefly described in this power point presentation .
The document discusses various methods for immobilizing microbial cells and enzymes, including carrier binding techniques like adsorption, covalent binding, cross-linking and entrapment. It describes common support materials like natural polymers, synthetic polymers and inorganic materials used for immobilization. The advantages of immobilization include recyclability, stability and potential applications in industries like food, biomedical and biodiesel production. Yeast cell immobilization using calcium alginate entrapment is provided as an example.
Cytotoxicity and genotoxicity of nanoparticleskumuthan MS
This document discusses the cytotoxicity and genotoxicity of nanoparticles. It outlines three main mechanisms by which nanoparticles can enter cells: direct diffusion across the cell membrane, endocytosis, and through membrane transporter proteins. The document then discusses various ways nanoparticles can be toxic to cells, such as releasing toxic ions, generating reactive oxygen species, or directly interacting with biological targets. In terms of genotoxicity, nanoparticles can directly interact with and damage DNA. They can also cause indirect genotoxicity by interacting with other nuclear proteins or disturbing cell processes, which can then lead to oxidative stress and DNA damage. The document concludes by noting nanoparticles may cause genotoxicity by inhibiting antioxidant defenses in cells.
A Biosensor is a device for the detection of an analyte that combines a biological component with a physio-chemical detector component.
Download: https://www.topicsforseminar.com/2014/10/biosensors-ppt.html
A biosensor is a device that integrates a biological component with a physicochemical detector. There are three main components: the biological recognition element, transducer, and associated electronics. The biological element interacts selectively with the analyte. The transducer converts this interaction into a quantifiable signal like a current or voltage. The associated electronics then process and display the results. Common types of biosensors include electrochemical, optical, and ion channel switch biosensors which detect analytes through electrochemical reactions, light interactions, or ion flow respectively.
This document provides an overview of biosensors, including their definition, components, principles of operation, examples, applications, and future potential. A biosensor integrates a biological recognition element with a physiochemical transducer to produce an electronic signal proportional to the concentration of an analyte. Common types include calorimetric, potentiometric, amperometric, and optical biosensors. Applications include medical diagnostics, environmental monitoring, food analysis, and industrial process control. The document concludes that biosensors can help identify materials and their concentrations in various fields.
This a short and efficient presentation On Biosensor for giving presentation in the upcoming seminar....
This could be more edited further for future purposes......
Contact: arnabguptakabiraj@gmail.com
This is for the beginners level giving presentation for the first time....
Protein based nanostructures for biomedical applications karoline Enoch
Proteins are kind of natural molecules that show unique
functionalities and properties in biological materials and
manufacturing feld. Tere are numerous nanomaterials
which are derived from protein, albumin, and gelatin. Tese
nanoparticles have promising properties like biodegradability, nonantigenicity, metabolizable, surface modifer, greater
stability during in vivo during storage, and being relatively
easy to prepare and monitor the size of the particles.
These particles have the ability to attach covalently with
drug and ligand
The document discusses structure-based drug design (SBDD). It first provides background on drug design and SBDD. It then describes some key aspects of SBDD, including using the 3D structure of the biological target obtained from techniques like X-ray crystallography and NMR spectroscopy. It also discusses ligand-based and receptor-based drug design approaches. The document then outlines the typical steps involved in SBDD, including target selection, ligand selection, target preparation, docking, evaluating results, and discusses some molecular docking techniques and scoring functions used to predict binding.
Aptamers are small nucleic acid molecules that bind target molecules with high affinity and specificity. They are generated through an in vitro selection process called SELEX and have several advantages over antibodies such as low immunogenicity and toxicity. Recent advancements have improved SELEX technology and clinical development of aptamer-based therapeutics is ongoing for cancer, eye diseases, and other applications. However, challenges remain around nuclease degradation, renal filtration, controlling duration of action, and potential toxicity that must be addressed for aptamers to reach their full potential as targeted therapeutics.
Enzyme immobilization ,Methods ,advantages and disadvantages and applicationsTaufica Nusrat
The document discusses immobilized enzymes, including definitions, important aspects of immobilization, and reasons for and limitations of immobilization. It describes the components of enzyme immobilization including enzymes, support matrices, and immobilization techniques. The document outlines properties required of support materials and classifications of supports as organic or inorganic. It details various immobilization techniques and provides examples of kinetics, factors affecting production, advantages/disadvantages, and applications of immobilized enzymes in food/beverage, pharmaceutical, and biomedical fields.
The document discusses aptamers, which are single-stranded folded oligonucleotides or peptides that bind to molecular targets with high affinity and specificity. Aptamers are produced through an in vitro selection process called SELEX that identifies nucleic acid sequences that bind to a target. The document outlines the SELEX process and compares properties of aptamers to antibodies. Potential applications of aptamers discussed include use as therapeutics, drug delivery agents, diagnostic tools, and in bioimaging and Western blot analysis due to their high specificity and low immunogenicity.
Biosensors: General Principles and ApplicationsBhatt Eshfaq
1. A biosensor is a device that uses specific biochemical reactions to detect chemical compounds in biological samples through the integration of a biological element with a physiochemical transducer.
2. Professor Leland C Clark Jr is considered the "Father of the Biosensor" for his work developing the first enzyme electrode for glucose detection in 1962.
3. There are various types of biosensors including calorimetric, potentiometric, amperometric, and optical biosensors that use different sensing techniques like fluorescence, DNA microarrays, and surface plasmon resonance.
Antisense therapy uses synthetic nucleic acid strands that bind to messenger RNA (mRNA) to inactivate genes and their protein production. It has potential applications for treating various diseases like cancer, diabetes, and genetic disorders. Antisense oligonucleotides work by binding to mRNA, preventing translation into protein via RNase H activity or steric hindrance. While promising, antisense drugs require further clinical trials to prove efficacy compared to other treatments.
Immobilization of enzyme and its applicationsKeshav Singh
IMMOBILIZATION OF ENZYME AND ITS APPLICATIONS
Need for Immobilization
Advantages of Immobilized Enzymes
Methods of immobilization
Carrier for immobilized enzyme
Applications of Immobilized Enzymes
The document provides an overview of protein-ligand docking, which is a computational method used in structure-based drug design to predict how small molecules bind to proteins. It discusses key components of docking software including search algorithms that generate poses of ligands in the binding site and scoring functions that calculate binding affinity scores. The document also touches on uses of docking like virtual screening and pose prediction, as well as considerations like flexible docking and handling protein conformations.
Nanoparticles show promise for biomedical imaging and diagnosis due to their large size and multifunctionality compared to small molecules. Magnetic iron oxide nanoparticles are commonly used in MRI because they shorten T2 relaxation times, allowing hydrogen protons to move closer to the magnet and produce clearer images. Various types of functionalized magnetic nanoparticles including amine, carboxyl, epoxy and IDA functionalized nanoparticles are used for applications like immunoassay, gene transfection, biomolecule separation, cell separation, enzyme immobilization, drug delivery, and biomedical imaging. Nanoparticles also show potential for targeted cancer drug delivery and simultaneous imaging and therapy.
TISSUE DEVELOPMENT WITH TISSUE ENGINEERING APPROACHFelix Obi
Tissue Engineering is the development and practice of combining scaffolds, cells, and suitable biochemical factors (regulatory factors or Signals) into functional tissues. The goal of tissue engineering is to assemble functional constructs that restore, maintain, or improve damaged tissues or whole organs.
Cells are the building blocks of tissue, and tissues are the basic unit of function in the body. Generally, groups of cells make and secrete their own support structures, called extracellular matrix. This matrix, or scaffold, does more than just support the cells; it also acts as a relay station for various signaling molecules. Thus, cells receive messages from many sources that become available from the local environment. Each signal can start a chain of responses that determine what happens to the cell. By understanding how individual cells respond to signals, interact with their environment, and organize into tissues and organisms, Tissue Engineers are now able to manipulate these processes to amend damaged tissues or even create new ones.
Characterization of the adhesive interactions between cells and biomaterialsDr. Sitansu Sekhar Nanda
This document discusses characterization of adhesive interactions between cells and biomaterials. It begins by introducing how biomaterials are used to facilitate cell adhesion during tissue repair or replacement. It then discusses various adhesion receptors in native tissue like integrins, their classification and role in connecting the extracellular and intracellular environments. The document also covers optimization of cellular adhesion through biomaterial modification and different methods to measure cell adhesion like micromanipulation, centrifugation and applying hydrodynamic shear stress. It concludes that modifying biomaterials to mimic native adhesive interactions can help control downstream cell responses and have therapeutic applications.
This document discusses affinity chromatography and biomimetic dyes. It begins by introducing affinity chromatography as a technique for separating substances based on their reversible interactions with immobilized ligands. It then provides historical background on affinity chromatography and describes various aspects of the technique including common matrices, ligand design and immobilization methods, and elution strategies. The document also discusses the use of biomimetic dyes as ligands for affinity chromatography, noting their low cost but lack of specificity. It describes strategies for designing new dye ligands with improved affinity and specificity for target proteins through mimicking natural ligands.
Application of Biological Assemblies in NanoBiotechnology PPtZohaib HUSSAIN
The document discusses several applications of biological assemblies in nanobiotechnology. It describes how biological assemblies, such as hemoglobin, are the functional forms of molecules and can be used to build larger structures. It then discusses using drug nanocrystals to improve drug solubility and bioavailability. Next, it covers using nano-containers like liposomes for targeted drug delivery. The document also discusses using protein crystals called S-layers for nanolithography and peptide templates for controlling biomineralization. Finally, it discusses the potential of utilizing biomineralization principles in nanotechnology applications like tissue engineering.
Immobilised enzymes ppt presentation - dr. r. mallikamallikaswathi
The document discusses immobilized enzymes and their applications. It defines immobilization as imprisoning cells or enzymes onto a support matrix. The first immobilized enzyme was amino acylase used in Japan. Immobilization offers benefits like enhanced efficiency, reproducibility, and stability. Common supports discussed include natural polymers like alginate and chitin, and inorganic materials like zeolites. The main immobilization methods covered are adsorption, covalent bonding, entrapment, and encapsulation. Important industrial uses of immobilized enzymes include production of antibiotics, beverages, amino acids, and their use in biomedical applications like disease diagnosis.
Biosensors are the analytical device that are used to measure the concentration of analye , these type of biosensors are made with conjugation of enzymes as a biological eliment to quantify a (bio)chemical substance / analyte are reffered to as Enzyme-probe Biosensors .
Biosensors are of many types but focusing on Enzyme biosensors there are 4 main types which are briefly described in this power point presentation .
The document discusses various methods for immobilizing microbial cells and enzymes, including carrier binding techniques like adsorption, covalent binding, cross-linking and entrapment. It describes common support materials like natural polymers, synthetic polymers and inorganic materials used for immobilization. The advantages of immobilization include recyclability, stability and potential applications in industries like food, biomedical and biodiesel production. Yeast cell immobilization using calcium alginate entrapment is provided as an example.
Cytotoxicity and genotoxicity of nanoparticleskumuthan MS
This document discusses the cytotoxicity and genotoxicity of nanoparticles. It outlines three main mechanisms by which nanoparticles can enter cells: direct diffusion across the cell membrane, endocytosis, and through membrane transporter proteins. The document then discusses various ways nanoparticles can be toxic to cells, such as releasing toxic ions, generating reactive oxygen species, or directly interacting with biological targets. In terms of genotoxicity, nanoparticles can directly interact with and damage DNA. They can also cause indirect genotoxicity by interacting with other nuclear proteins or disturbing cell processes, which can then lead to oxidative stress and DNA damage. The document concludes by noting nanoparticles may cause genotoxicity by inhibiting antioxidant defenses in cells.
A Biosensor is a device for the detection of an analyte that combines a biological component with a physio-chemical detector component.
Download: https://www.topicsforseminar.com/2014/10/biosensors-ppt.html
A biosensor is a device that integrates a biological component with a physicochemical detector. There are three main components: the biological recognition element, transducer, and associated electronics. The biological element interacts selectively with the analyte. The transducer converts this interaction into a quantifiable signal like a current or voltage. The associated electronics then process and display the results. Common types of biosensors include electrochemical, optical, and ion channel switch biosensors which detect analytes through electrochemical reactions, light interactions, or ion flow respectively.
This document provides an overview of biosensors, including their definition, components, principles of operation, examples, applications, and future potential. A biosensor integrates a biological recognition element with a physiochemical transducer to produce an electronic signal proportional to the concentration of an analyte. Common types include calorimetric, potentiometric, amperometric, and optical biosensors. Applications include medical diagnostics, environmental monitoring, food analysis, and industrial process control. The document concludes that biosensors can help identify materials and their concentrations in various fields.
This a short and efficient presentation On Biosensor for giving presentation in the upcoming seminar....
This could be more edited further for future purposes......
Contact: arnabguptakabiraj@gmail.com
This is for the beginners level giving presentation for the first time....
Protein based nanostructures for biomedical applications karoline Enoch
Proteins are kind of natural molecules that show unique
functionalities and properties in biological materials and
manufacturing feld. Tere are numerous nanomaterials
which are derived from protein, albumin, and gelatin. Tese
nanoparticles have promising properties like biodegradability, nonantigenicity, metabolizable, surface modifer, greater
stability during in vivo during storage, and being relatively
easy to prepare and monitor the size of the particles.
These particles have the ability to attach covalently with
drug and ligand
The document discusses structure-based drug design (SBDD). It first provides background on drug design and SBDD. It then describes some key aspects of SBDD, including using the 3D structure of the biological target obtained from techniques like X-ray crystallography and NMR spectroscopy. It also discusses ligand-based and receptor-based drug design approaches. The document then outlines the typical steps involved in SBDD, including target selection, ligand selection, target preparation, docking, evaluating results, and discusses some molecular docking techniques and scoring functions used to predict binding.
Aptamers are small nucleic acid molecules that bind target molecules with high affinity and specificity. They are generated through an in vitro selection process called SELEX and have several advantages over antibodies such as low immunogenicity and toxicity. Recent advancements have improved SELEX technology and clinical development of aptamer-based therapeutics is ongoing for cancer, eye diseases, and other applications. However, challenges remain around nuclease degradation, renal filtration, controlling duration of action, and potential toxicity that must be addressed for aptamers to reach their full potential as targeted therapeutics.
Enzyme immobilization ,Methods ,advantages and disadvantages and applicationsTaufica Nusrat
The document discusses immobilized enzymes, including definitions, important aspects of immobilization, and reasons for and limitations of immobilization. It describes the components of enzyme immobilization including enzymes, support matrices, and immobilization techniques. The document outlines properties required of support materials and classifications of supports as organic or inorganic. It details various immobilization techniques and provides examples of kinetics, factors affecting production, advantages/disadvantages, and applications of immobilized enzymes in food/beverage, pharmaceutical, and biomedical fields.
The document discusses aptamers, which are single-stranded folded oligonucleotides or peptides that bind to molecular targets with high affinity and specificity. Aptamers are produced through an in vitro selection process called SELEX that identifies nucleic acid sequences that bind to a target. The document outlines the SELEX process and compares properties of aptamers to antibodies. Potential applications of aptamers discussed include use as therapeutics, drug delivery agents, diagnostic tools, and in bioimaging and Western blot analysis due to their high specificity and low immunogenicity.
Biosensors: General Principles and ApplicationsBhatt Eshfaq
1. A biosensor is a device that uses specific biochemical reactions to detect chemical compounds in biological samples through the integration of a biological element with a physiochemical transducer.
2. Professor Leland C Clark Jr is considered the "Father of the Biosensor" for his work developing the first enzyme electrode for glucose detection in 1962.
3. There are various types of biosensors including calorimetric, potentiometric, amperometric, and optical biosensors that use different sensing techniques like fluorescence, DNA microarrays, and surface plasmon resonance.
Antisense therapy uses synthetic nucleic acid strands that bind to messenger RNA (mRNA) to inactivate genes and their protein production. It has potential applications for treating various diseases like cancer, diabetes, and genetic disorders. Antisense oligonucleotides work by binding to mRNA, preventing translation into protein via RNase H activity or steric hindrance. While promising, antisense drugs require further clinical trials to prove efficacy compared to other treatments.
Immobilization of enzyme and its applicationsKeshav Singh
IMMOBILIZATION OF ENZYME AND ITS APPLICATIONS
Need for Immobilization
Advantages of Immobilized Enzymes
Methods of immobilization
Carrier for immobilized enzyme
Applications of Immobilized Enzymes
The document provides an overview of protein-ligand docking, which is a computational method used in structure-based drug design to predict how small molecules bind to proteins. It discusses key components of docking software including search algorithms that generate poses of ligands in the binding site and scoring functions that calculate binding affinity scores. The document also touches on uses of docking like virtual screening and pose prediction, as well as considerations like flexible docking and handling protein conformations.
Nanoparticles show promise for biomedical imaging and diagnosis due to their large size and multifunctionality compared to small molecules. Magnetic iron oxide nanoparticles are commonly used in MRI because they shorten T2 relaxation times, allowing hydrogen protons to move closer to the magnet and produce clearer images. Various types of functionalized magnetic nanoparticles including amine, carboxyl, epoxy and IDA functionalized nanoparticles are used for applications like immunoassay, gene transfection, biomolecule separation, cell separation, enzyme immobilization, drug delivery, and biomedical imaging. Nanoparticles also show potential for targeted cancer drug delivery and simultaneous imaging and therapy.
TISSUE DEVELOPMENT WITH TISSUE ENGINEERING APPROACHFelix Obi
Tissue Engineering is the development and practice of combining scaffolds, cells, and suitable biochemical factors (regulatory factors or Signals) into functional tissues. The goal of tissue engineering is to assemble functional constructs that restore, maintain, or improve damaged tissues or whole organs.
Cells are the building blocks of tissue, and tissues are the basic unit of function in the body. Generally, groups of cells make and secrete their own support structures, called extracellular matrix. This matrix, or scaffold, does more than just support the cells; it also acts as a relay station for various signaling molecules. Thus, cells receive messages from many sources that become available from the local environment. Each signal can start a chain of responses that determine what happens to the cell. By understanding how individual cells respond to signals, interact with their environment, and organize into tissues and organisms, Tissue Engineers are now able to manipulate these processes to amend damaged tissues or even create new ones.
Characterization of the adhesive interactions between cells and biomaterialsDr. Sitansu Sekhar Nanda
This document discusses characterization of adhesive interactions between cells and biomaterials. It begins by introducing how biomaterials are used to facilitate cell adhesion during tissue repair or replacement. It then discusses various adhesion receptors in native tissue like integrins, their classification and role in connecting the extracellular and intracellular environments. The document also covers optimization of cellular adhesion through biomaterial modification and different methods to measure cell adhesion like micromanipulation, centrifugation and applying hydrodynamic shear stress. It concludes that modifying biomaterials to mimic native adhesive interactions can help control downstream cell responses and have therapeutic applications.
This document discusses affinity chromatography and biomimetic dyes. It begins by introducing affinity chromatography as a technique for separating substances based on their reversible interactions with immobilized ligands. It then provides historical background on affinity chromatography and describes various aspects of the technique including common matrices, ligand design and immobilization methods, and elution strategies. The document also discusses the use of biomimetic dyes as ligands for affinity chromatography, noting their low cost but lack of specificity. It describes strategies for designing new dye ligands with improved affinity and specificity for target proteins through mimicking natural ligands.
Effect of Surface Engineering on Stem Cells.pdfaman15nanavaty
A thorough review covering the nascent domain of Surface Engineering on Stem Cells. This short review will cover basic details of Stem Cell Engineering and Cell Culture Surface Engineering.
This document describes a study on the immobilization of the enzyme β-galactosidase using plasma-deposited organosilicon thin films. The study developed a two-step procedure where β-galactosidase was first adsorbed onto a substrate, then overcoated with a porous plasma polymerized 1,1,3,3-tetramethyldisiloxane film. This entrapped the enzymes while allowing substrate molecules to penetrate and be hydrolyzed. The procedure produced a biofunctional coating where β-galactosidase was entrapped in the polymer matrix while preserving its structure and activity. This dry process using plasma technology provides a fast, simple, and mild method for enzyme
Affinity chromatography is a technique used to separate biochemical compounds based on a reversible interaction between a compound and a ligand coupled to a chromatography matrix. It offers selectivity and can purify compounds that may be difficult to separate by other techniques. Key aspects of affinity chromatography include the matrix, ligand, ligand immobilization through various coupling methods, and elution techniques to reverse binding. Biomimetic dyes designed to mimic natural ligands can function as ligands in affinity chromatography.
Nanomediated anticancer drug delivery.pptxMsRicha2
Nanoparticles have potential for targeted anticancer drug delivery. They can be engineered to actively or passively target tumors. Passive targeting relies on nanoparticles' ability to accumulate in tumors through the enhanced permeability and retention effect, which is caused by tumors' leaky blood vessels and poor lymphatic drainage. Nanoparticles can also be engineered for active targeting by attaching ligands that bind to receptors on cancer cells. The small size of nanoparticles allows them to penetrate tissues and cell membranes more readily than traditional drugs to selectively deliver anticancer therapies and improve treatment outcomes with fewer side effects.
Plant cell immobilization techniques confine catalytically active plant cells within a reactor system to increase productivity. Common techniques include entrapment within a polymer matrix, microencapsulation, and adsorption to a surface. Immobilized cells are protected from shear forces and can operate continuously in bioreactors. While immobilization allows high biomass levels and simplified downstream processing, it may reduce cell biosynthesis capacity and require the release of products from the immobilized cells.
Discussing advances in Magnetic Bead coating technologies - Page 9 & 10 - Article from Joshua Soldo from Australian listed Biotech company Anteo Diagnostics ASX:ADO
Stem cells and nanotechnology in regenerative medicine and tissue engineeringDr. Sitansu Sekhar Nanda
Alexis Carrel, winner of the Nobel Prize in Physiology or Medicine in 1912 and the father of whole-organ transplant, was the first to develop a successful technique for end to end arteriovenous anastomosis in transplantation.
Nanogels are particles composed of physically or chemically cross linked polymer networks that expand in an appropriate solvent. Nanogels are hydrophilic three dimensional networks. Due to their relatively high drug encapsulation ability, consistency, tunable size, effortless preparation, negligible toxicity, and stability in the presence of serum, including stimuli responsiveness, these studies integrate characteristics for topical drug delivery. These are soluble in water and permit immediate drug loading in aqueous media. These are created using a vast array of methods, including photolithographic technique, membrane emulsification, and polymerization methods. Due to the entrapment of nanoparticles in the gel matrix, nanogels used as dermatological preparations have prolonged exposure times on the skin, thereby extending the duration of therapeutic efficacy. B. Karthikeyan | G. Alagumanivasagam "A Review on Nanogels" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-7 | Issue-3 , June 2023, URL: https://www.ijtsrd.com.com/papers/ijtsrd57514.pdf Paper URL: https://www.ijtsrd.com.com/pharmacy/other/57514/a-review-on-nanogels/b-karthikeyan
This document provides an overview of regenerative periodontal surgery techniques. It discusses the historical concepts of periodontal regeneration including bone grafts, guided tissue regeneration (GTR), and the emerging field of tissue engineering. Key cellular mediators and signaling molecules that can promote periodontal regeneration are described, including platelet-derived growth factor, bone morphogenetic proteins, insulin-like growth factor, and enamel matrix derivative. The document also reviews the different cell types involved in periodontal regeneration, including dental pulp stem cells, periodontal ligament stem cells, dental follicle progenitor cells, and dental epithelial stem cells. The criteria for achieving true periodontal regeneration and methods to guide cell differentiation and maturation are also summarized.
1) The document discusses various barriers to targeting tumors including heterogeneity in blood flow within tumors and overexpression of efflux transporters in tumor cells.
2) It describes three main approaches to overcoming these barriers: passive targeting using the EPR effect, active targeting by attaching targeting ligands like antibodies, and physical targeting using stimuli like pH, temperature, or magnetic fields.
3) Examples are given of using each approach, such as pH-sensitive nanoparticles that degrade in the acidic tumor environment or magnetic drug targeting using nanoparticles guided by an external magnet.
Tissue engineering uses scaffolds, cells, and signaling molecules to regenerate tissues and organs. Scaffolds provide a structure for cell attachment, growth, and tissue formation. Natural polymers like collagen and hyaluronic acid, and synthetic polymers like poly-lactic-co-glycolic acid are commonly used as scaffold materials. Scaffolds can be fabricated using various methods including freeze drying, electrospinning, 3D printing, and textile technologies to produce scaffolds with desirable properties like porosity and pore size for tissue growth. Scaffolds seeded with stem cells or tissue-specific cells aim to repair and regenerate tissues for applications in skin, bone, cartilage, and other organs.
Methods to Improve Biocompatibility of MaterialsPeeyush Mishra
Methods to improve the biocompatibility of materials include surface coating, surface patterning, and zwitterionic surface coating. Surface coating involves using electrically neutral and hydrophilic polymers like PEG and PLGA. Surface patterning reduces the diffusive transport zone through nanostructured surfaces. Zwitterionic coatings use materials with equal positive and negative charges like polyampholytes to create an ultralow fouling surface. Future work aims to develop smaller biomolecule coatings like amino acids.
This document summarizes research on modifying bacterial cellulose through the addition of hyaluronic acid and gelatin during fermentation to produce novel nanocomposites for use as biomaterials in stem cell therapy. Transmission infrared spectroscopy and scanning electron microscopy showed the influence of hyaluronic acid and gelatin on the bacterial cellulose structure. Cell viability studies with human dental pulp stem cells demonstrated higher cell adhesion to bacterial cellulose scaffolds containing hyaluronic acid or gelatin over time. Confocal microscopy confirmed cell adhesion and distribution within the scaffold fibers. This research presents a new approach for developing natural nanocomposites using bacterial cellulose as scaffolds for regenerative medicine applications.
JBEI Research Highlight Slides - August 2022SaraHarmon4
The document discusses engineering yeast to produce the anti-cancer drug vinblastine through a long biosynthetic pathway involving 30 enzymatic steps from multiple plants. This provides an alternative microbial supply chain that does not rely on low-yielding extraction from plants. The engineered yeast is able to produce the vinblastine precursors catharanthine and vindoline, demonstrating the potential for sustainable production of complex plant metabolites through synthetic biology.
Layer by layer edible coating on fruits and vegetablesindu indu
This document presents information on layer by layer (LbL) edible coatings for fruits and vegetables. It discusses what edible coatings are, how the LbL technique works using alternating layers of positively and negatively charged materials, and examples of raw materials used like chitosan, gluten and whey protein. A case study is described where different polysaccharide coatings were tested on mandarins to enhance quality and storage time, including combinations of carboxymethyl cellulose, chitosan and fatty acids. Evaluation methods for coated fruits included firmness, weight loss, gas composition and sensory properties.
This document provides an overview of nanogels for drug delivery applications. It defines nanogels as nanosized polymer networks that swell in solvent. Nanogels have properties like biocompatibility and drug loading capacity. They can be administered via various routes and classified based on responsive behavior or linkage type. The document discusses synthesis, characterization, and applications of nanogels in cancer treatment, ophthalmic use, and more. Nanogels are a promising drug delivery system due to abilities like controlled drug release and delivery of therapeutics to targeted sites.
Recent Advancement and Patents of the Lipid Polymer Hybrid Nanoparticlespeertechzpublication
In recent years, robustness and surface engineering of dosage form made improvement in
pharmacokinetics with decrease in dose of drug. Specifi city with adherence of ligands has now become
the reality as surface modifi cation can easily deceive phagocytic system. Lipid molecules ensures the
release of drug at lymphatic system, entrapment of polymeric nanoparticles in lipoidal core led to the
avoidance of disadvantage of low entrapment effi ciency if use of hydrophobic drug with hydrophobic
polymer becomes essential. Various studies have been published and the best formulations with optimal
In vitro and In vivo results are highlighted in this paper. In this review most advanced researches and
accepted patents were discussed so to act as a medium for getting everything regarding lipid polymer
hybrid particles under one umbrella.
In recent years, robustness and surface engineering of dosage form made improvement in pharmacokinetics with decrease in dose of drug. Specificity with adherence of ligands has now become the reality as surface modifi cation can easily deceive phagocytic system. Lipid molecules ensures the
release of drug at lymphatic system, entrapment of polymeric nanoparticles in lipoidal core led to the
avoidance of disadvantage of low entrapment effi ciency if use of hydrophobic drug with hydrophobic polymer becomes essential. Various studies have been published and the best formulations with optimal In vitro and In vivo results are highlighted in this paper. In this review most advanced researches and accepted patents were discussed so to act as a medium for getting everything regarding lipid polymer hybrid particles under one umbrella.
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
NVBDCP.pptx Nation vector borne disease control programSapna Thakur
NVBDCP was launched in 2003-2004 . Vector-Borne Disease: Disease that results from an infection transmitted to humans and other animals by blood-feeding arthropods, such as mosquitoes, ticks, and fleas. Examples of vector-borne diseases include Dengue fever, West Nile Virus, Lyme disease, and malaria.
Knee anatomy and clinical tests 2024.pdfvimalpl1234
This includes all relevant anatomy and clinical tests compiled from standard textbooks, Campbell,netter etc..It is comprehensive and best suited for orthopaedicians and orthopaedic residents.
share - Lions, tigers, AI and health misinformation, oh my!.pptxTina Purnat
• Pitfalls and pivots needed to use AI effectively in public health
• Evidence-based strategies to address health misinformation effectively
• Building trust with communities online and offline
• Equipping health professionals to address questions, concerns and health misinformation
• Assessing risk and mitigating harm from adverse health narratives in communities, health workforce and health system
These lecture slides, by Dr Sidra Arshad, offer a quick overview of the physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar lead (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
6. Describe the flow of current around the heart during the cardiac cycle
7. Discuss the placement and polarity of the leads of electrocardiograph
8. Describe the normal electrocardiograms recorded from the limb leads and explain the physiological basis of the different records that are obtained
9. Define mean electrical vector (axis) of the heart and give the normal range
10. Define the mean QRS vector
11. Describe the axes of leads (hexagonal reference system)
12. Comprehend the vectorial analysis of the normal ECG
13. Determine the mean electrical axis of the ventricular QRS and appreciate the mean axis deviation
14. Explain the concepts of current of injury, J point, and their significance
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. Chapter 3, Cardiology Explained, https://www.ncbi.nlm.nih.gov/books/NBK2214/
7. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
1. Immobilization of
biomolecules on the
surface of biomaterials
By: Mohsen Norouzi
MSc Student of Tissue Engineering
Islamic Azad University of Najafabad (IAUN)
1
2. Biomaterials must possess bulk properties that
permit its function in the bio‐environment, but
also the best surface properties.
It is difficult to design materials that fulfill both
needs.
A common approach is to fabricate with
adequate bulk properties followed by a
special treatment to enhance the surface
properties.
Preface
2
3. Preface
The broad interdisciplinary area where properties and
processes at this interface are investigated and biofunctional
surfaces are fabricated is called Biological Surface Science.
Examples:
medical implants in the human body (dental implants, artificial
hip and knee joints, artificial blood vessels and heart valves, etc.)
tissue engineering
biosensors and biochips for diagnosis (DNA‐chips, etc.)(clinical
diagnostics, environmental control, food production)
Bioelectronics (systems to get information storage and processing
) and artificial photosynthesis (clean energy)
biomimetic materials (mimic the functional properties of
biological materials/components in order to achieve new and
better materials; ow friction from the sharkskin or self‐cleaning
character like the lotus leaf )
3
5. Approaches to improve biointerfaces:
reduction of unspecific protein adsorption
enhanced adsorption of specific proteins
material modification by immobilization of cell
recognition motives to obtain controlled
interaction between cells and synthetic substrate
• Using methods like selfassembly (SAMs),
surface modification, photochemical
immobilization or polymer chemistry,
complex surfaces with immobilized
peptides and proteins can be prepared
Preface
5
6. Biomolecules used in precision immobilization
strategies include proteins, lipids, polypeptides,
polynucleotides and polysaccharides
Immobilization techniques range from relatively
low to extremely high specificity.
characteristics of successful precision
engineered biorecognition surfaces:
presence of one ligand site and the receptor‐ligand
affinity
an appropriate surface density of those sites
spatial distribution of the ligands
Preface
6
7. The use of short peptides for surface
biorecognition has proved to be
advantageous over the use of the long
chain native ECM proteins, since the
latter tend to be randomly folded upon
adsorption, being the receptor binding
domains not always sterically
available.
Preface
7
8. Immobilization
Molecules may be immobilized either passively through;
o Hydrophobic
o Ionic interactions
o Covalently by attachment to activated surface groups.
Non-covalent surfaces are effective for many applications;
however, passive adsorption fails in many cases.
Covalent immobilization is often necessary for binding of
molecules that do not adsorb, adsorb very weakly, or adsorb
with improper orientation and conformation to non-covalent
surfaces.
Covalent immobilization may result in better biomolecule
activity, reduced nonspecific adsorption, and greater stability.
8
9. Immobilization
Immobilization reaction should have
several characteristics;
Firstly, the reaction should occur rapidly and
therefore allow the use of low concentrations of
reagents for immobilization.
The chemistry should require little, if any, post-
synthetic modification of ligands before
immobilization to maximize the number of
compounds that can be generated by solution
or solid-phase synthesis and minimize the cost
of these reagents.
Immobilized ligands must be in an oriented
and homogeneous manner.
9
10. Immobilization
The immobilization process should occur
selectively in the presence of common
functional groups, including amines, thiols,
carboxylic acids, and alcohols.
Amino-NH2,
Carboxy-COOH,
Aldehyde-CHO,
Thiol-SH,
Hydroxyl-OH
10
11. Immobilization
Surface density of the ligand should be
optimized.
Low density surface coverage will yield a
correspondingly low frequency.
High surface densities may result steric
interference between the covalently
immobilized ligand molecules, impending
access to the target molecules.
11
12. 1) unhindered binding. 2) inaccessible binding site. 3) hindered
binding site when adjacent site is occupied. 4) restricted access
binding site.
Immobilization
12
13. Immobilization
Correct orientation of the ligand molecules on the surface, and using
a spacer arm are important and critical and makes the ligand
available for the target.
13
14. Proteins are much more sensitive
to their physiological environments
and can easily be degraded or
denaturated by physical or
chemical effects. Protein`s 3-D
confirmation must not change
during immobilization procedure.
DNA molecules are much more
stable then proteins.
It is easier to immobilize DNA
molecules.
Immobilization
14
15. Preparation of Surface for
Biomolecule Immobilization
Modification of the surface to create
functional groups.
Modification of biomolecules for
covalent attachment to the surface.
15
21. Surface engineered scaffolds
Collagen:
major structural component forming the natural ECM of
connective tissues and organs
one of the most established methods for endowing cell
adhesive properties to the scaffolds
Examples:
PLA and PLGA scaffolds chemically grafted with collagen by
plasma treatment have shown enhanced adhesion and
spreading of fibroblasts
Collagen modification by conjugation reactions onto PLA
scaffolds grafted with polymethacrylic acid also has improved
cell spreading and growth for use in cartilage tissue
engineering.
its immunogenicity has limited its applications
21
22. Gelatin:
a good alternative for collagen because of its
absence of antigenicity and ease of handling
at high concentrations
Example:
Gelatin immobilized onto porous scaffolds by
physical entrapment and chemical crosslinking
showed greatly enhanced surface properties on
attachment, proliferation, and ECM deposition of
osteoblasts
Surface engineered scaffolds
22
23. Cell adhesive peptides:
Rather than immobilizing the whole protein, chemical
conjugation of short chain peptide moieties derived from
the cell adhesive proteins onto the polymer surface can be
a much more effective strategy
Advantages of The surface immobilization of short peptides:
higher stability against conformational change
easy controllability of surface density,
orientation more favorable for ligand–receptor interaction and
cell adhesion
minimizing immune responses and infection
Surface engineered scaffolds
23
24. peptide sequences involved in cellular
interactions by receptor binding:
RGD, IKVAV, and YIGSR
RGD sequence: one of the best known foruse in tissue
engineering applications
Examples:
Immobilization of RGD onto 3-D matrices to improve cell
adhesive properties was previously demonstrated in collagen
gels, showing enhanced adherence of murine melanoma cells
RGD, along with other short peptide sequences such as IKVAV,
YIGSR, RNAIAEIIKDI from laminin, and HAV from N-cadherin, was
also used for engineering of neural tissue.
PLA scaffolds modified with RGD by plasma treatment not only
resulted in improved adhesion of the osteoblast-like cells, but
also supported its growth and differentiation
osteoblasts seeded onto the RGD immobilized scaffolds greatly
enhanced mineralization and formation of bone-like tissues
24
26. Hyaluronic acid:
a non-sulfated glycosaminoglycan (GAG), is
a major substance of the gel-like component
in the extracellular matrix of connective
tissues
capable of specific cell interaction via the
CD44 receptor which promotes wound
healing and induces chondrogenesis
Examples:
Chitosan–gelatin composite scaffolds modified with
HA have been shown to increase the adhesion of
fibroblasts
PLGA scaffolds modified with HA supported the
growth of chondrocytes with maintenance of its
original phenotype, showing great potential for
cartilage tissue engineering
26
27. Galactose:
utilized in scaffolds for liver tissue engineering
recognized by mammalian hepatocytes through
the asialoglyco protein receptor leading to
regulation of a degradative pathway I
glycoprotein homeostasis
Examples:
Porous scaffolds immobilized with galactose have been
fabricated to improve hepatocyte attachment, viability,
and metabolic functions. Gelatin sponges modified with
galactose were shown to support hepatocyte adhesion
and function such as release of lactate dehydrogenase
(LDH), albumin secretion, and urea synthesis. Perfusion
culture of hepatocytes with galactose-derivatized PLGA
scaffolds further improved viability and functional
activity of the cells
27
28. Heparin:
intensively studied for growth factor releasing
matrices in tissue engineering.
a highly sulfated GAG constituting the
extracellular matrix, and is known for its specific
interactions with various angiogenic growth
factors
Examples:
Heparin binding has been shown to preserve the
stability and biological activity of the growth factors. A
wide variety of scaffold matrices, including nanofibers,
prepared from collagen, fibrin, chitosan, alginate, PLA
and PLGA, have been incorporated or immobilized with
heparin to achieve sustained release of growth factors
28
38. Basement Material (Substrate):
Synthetic polymer substrates, polystyrene (PS) and poly(lactic-co-
glycolic acid) (PLGA), polydimethylsiloxane (PDMS), silica (Si) and
titanium (Ti).
Linkage Material:
Polydopamine
Chemical/Physical Method:
Dipcoating a biomimetic polymer (PD) thin film onto the polymer
surface followed by conjugation of adhesion peptides and
neurotrophic growth factors to the biomimetic polymer film.
Because amine and thiol groups can be covalently conjugated to a
PD layer via the quinone group, PD coating exhibits latent reactivity
to various nucleophiles with those functional groups
Immobilized Material:
ECM protein-derived adhesion peptides, fibronectin [Arg-Gly-Asp
(RGD)] and laminin [Try-Ile-Gly-Ser-Arg (YIGSR)], and neurotrophic
factors, NGF and GDNF
Goal:
Modification of tissue engineering scaffolds for improving the
efficacy of stem cell therapy by generating physicochemical
stimulation promoting proliferation and differentiation of stem cells
surface modification for efficient and reliable manipulation of
human neural stem cell (NSC) differentiation and proliferation
38
39. Result/Effectiveness:
highly efficient, simple immobilization of neuro trophic growth factors
and adhesion peptides onto polymer substrates.
greatly enhance differentiation and proliferation of human NSCs
(human fetal brain derived NSCs and human induced pluripotent stem
cell derived NSCs) at a level comparable or greater than currently
available animal derived coating materials (Matrigel) with safety issues.
versatile platform technology for developing chemically defined, safe,
functional substrates and scaffolds for therapeutic applications of
human NSCs.
efficient surface immobilization of proteins and peptides to a diverse
range of materials, including polymer scaffolds, ceramic substrates, and
metal devices, for stem cell culture and transplantation.
versatile platform technology for efficient development of biomimetic
substrates and scaffolds that induce desirable stem cell behavior and
enhance stem cell function
39
42. Basement Material (Substrate):
ZrO2, TiZr and Ti with its naturally occurring oxide layer TiO2
Linkage Material:
Specific adsorbing peptides (Pep5 (SHKHGGHKHGGH KHGSSGKG)) are
used as anchor molecules to immobilize oligodesoxynucleotides (ODNs)
on the implant surface (anchor strand, AS)
Chemical/Physical Method:
The BAM is conjugated to a complementary ODN strand (CS) which is
able to hybridize to the AS on the implant surface to immobilize the
BAM. The ODN double strand allows for a controlled release of the BAM
adjustable by the ODN sequence and length.
Immobilized Material:
biologically active molecules (BAMs), e.g. antibiotics or growth
factors immobilize the parathyroid hormone (PTH) fragment 1-34
42
43. Result/Effectiveness:
Successful immobilization of biologically active PTH (1-34)
The high potential of the established surfaces to achieve an increased
osseointegration of variable implants, especially for patients with risk
factors. the development of bioinductive implant surfaces might
increase the healing capacity in the bone, especially for patients with
risk factors such as osteoporosis, where the healing of bone fractures is
disturbed.
The ability of PTH (1-34) to induce the differentiation of osteoblast
precursor cells C2C12 was detected by the quantification of the ALP
activity.
The conjugation of PTH with CS only slightly decreased the Alkaline
phosphatase(ALP) activity, indicating that the biological activity was
almost completely maintained. The application of the immobilization
system on the three materials allows for the modification of the surfaces
with PTH (1-34) as the ALP activity could be increased while
unspecifically bound PTH (1-34) itself showed no effect.
43
46. Basement Material (Substrate):
gold, platinum, glass and titanium
Linkage Material:
Peptide motifs
Chemical/Physical Method:
We synthesized bifunctional quartz-binding peptide QBP1–RGD and
titanium-binding peptide TiBP1–RGD peptides via solid phase peptide
synthesis and immobilizes these peptide conjugates on the surface through
directed assembly in a single step
Immobilized Material:
poly(ethylene glycol) anti-fouling polymer and the integrin-binding
RGD sequence
46
47. Result/Effectiveness:
We successfully imparted cell-resistant properties to gold and platinum
surfaces using gold- and platinum-binding peptides, respectively, in
conjunction with PEG.
several-fold increase in the number and spreading of fibroblast cells on
glass and titanium surfaces using quartz and titanium-binding peptides
in conjunction with the integrin ligand RGD.
Control over the extent of cell–material interactions by relatively simple
and biocompatible surface modification procedures using inorganic
binding peptides as linker molecules.
Targeted assembly proved to be an efficient way of immobilizing large
molecules (i.e. PEG) through, first, coating the inorganic binding
peptides and then performing the conjugation reaction.
Directed assembly, on the other hand, is preferred for the immobilization
of small molecules by synthesizing a single chimeric molecule with bi
functional domains.
Control over the extent of cell–material interactions can be achieved
by relatively simple and biocompatible surface modification procedures
using GEPIs as linker molecules.
QBP1 and the TiBP1 facilitate the immobilization of RGD on both
surfaces while preserving its functionality as a recognition site for cells.
47
50. References:
Prof. Marco Mascini, Immobilization of Biomollecules, Grenoble,
2004.
Laia Francesch de Castro , Surface modification of polymers by
plasma polymerization techniques for tissue engineering, doctorate
thesis, Universitat liull, Barcelona .
Hyun Jung Chung, Tae Gwan Park, Surface engineered and drug
releasing pre-fabricated scaffolds for tissue engineering, Advanced
Drug Delivery Reviews 59 (2007) 249–262.
Tina Micksch et al, A modular peptide-based immobilization system
for ZrO2, TiZr and TiO2 surfaces, Acta Biomaterialia (2014).
Qian Yu et al, Anti-fouling bioactive surfaces, Acta Biomaterialia 7
(2011) 1550–1557.
Dmitriy Khatayevich et al, Biofunctionalization of materials for
implants using engineered peptides, Acta Biomaterialia 6 (2010)
4634–4641.
Kisuk Yang et al, Polydopamine-mediated surface modification of
scaffold materials for human neural stem cell engineering,
Biomaterials 33 (2012) 6952e6964.
50