SPR and Avidin-Biotin interaction notes.pptx
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Surface plasmon resonance (SPR) biosensors detect analytes by measuring changes in refractive index when the analyte binds to a recognition element on the sensor surface. SPR biosensing utilizes the extremely high affinity interaction between avidin and biotin, with one of the partners immobilized on the sensor and the other used to attach the analyte. The avidin-biotin interaction provides a robust and stable link with affinity over 10^15 M-1 and has widespread applications in biosensors and other areas due to its specificity, strength, and ability to amplify signals.
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Here are the key differences between avidin, streptavidin, CaptAvidin and neutravidin:
- Avidin and streptavidin are both tetrameric proteins composed of four subunits, each capable of binding one biotin molecule.
- Streptavidin is derived from Streptomyces avidinii while avidin is derived from egg whites.
- Streptavidin is more resistant to denaturation than avidin.
- CaptAvidin is an avidin derivative where the biotin binding can be reversed by raising the pH, allowing isolation of biotinylated molecules.
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Here are the key differences between avidin, streptavidin, CaptAvidin and neutravidin:
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- Streptavidin is derived from Streptomyces avidinii while avidin is derived from egg whites.
- Streptavidin is more resistant to denaturation than avidin.
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This document discusses various electrochemical methods and their applications. It covers topics like potentiometry, types of electrochemical cells, reference electrodes, quantitative applications in clinical and environmental analysis, and potentiometric titrations. Ion selective electrodes are explained in detail, including how they work, types of analytes and electrodes, maintenance, and applications in various fields like analytical chemistry, clinical diagnostics, and environmental monitoring. Different types of voltammetry techniques are also introduced.
DNA Replication in eukaryotic and prokryotes
DNA replication involves the duplication of DNA prior to cell division. It occurs during the S phase of interphase in the cell cycle. There are three main modes of DNA replication: semiconservative, conservative, and dispersive. Semiconservative replication is the mechanism that occurs in eukaryotic cells, where the parental DNA strands separate and each acts as a template for new strand synthesis. This results in one old and one new strand in each of the two daughter DNA molecules after replication.
Analytical tool in Biochemistry and chemistry
The document discusses centrifugation, which uses centrifugal force to separate particles in a liquid based on density. It can be used to separate blood components. A centrifuge spins a sample at high speeds using rotors to generate centrifugal force much stronger than gravity. Denser particles sediment faster and settle at the bottom. Centrifugation is used in laboratories and industries for separation, purification, and analysis of substances.
xenobiotics chemistry and drug metabolism
Xenobiotics are compounds foreign to the body that are subject to biotransformation. Biotransformation involves two phases: Phase I uses enzymes like cytochrome P450 to introduce functional groups, making compounds slightly more hydrophilic. Phase II involves conjugating compounds with endogenous molecules like glutathione and glucuronic acid, greatly increasing hydrophilicity and facilitating excretion. Biotransformation can activate or inactivate compounds, and is influenced by genetic and environmental factors. The liver is the primary site of biotransformation, though other tissues play roles.
Disorders of Calcium and Phosphate Metabolism.ppt
This document discusses disorders of calcium and phosphate metabolism. It begins by explaining how vitamin D regulates calcium and phosphate homeostasis by increasing absorption in the gut and mobilization from bone, while stimulating calcium reabsorption and phosphate excretion in the kidney. Major disorders mentioned include chronic inflammatory conditions, infections, and cancers. The major mediators of calcium and phosphate balance, parathyroid hormone and calcitriol, are also described. The document provides overviews of calcium and phosphate biochemical metabolism and homeostasis, outlining causes of hypercalcemia and hyperphosphatemia.
SBC_450_Hereditary eye dysfunction.pptx 1.ppt
This document discusses several common eye conditions: cataracts, diabetic retinopathy, glaucoma, and age-related macular degeneration. It provides details on the signs and symptoms, causes, management, and treatment options for cataracts and diabetic retinopathy. Specifically, it explains that cataracts are cloudy areas in the lens of the eye that can impair vision. The most common treatment is cataract surgery. It also outlines the pathogenesis and classification of diabetic retinopathy, a complication of diabetes that damages the retina and can potentially lead to vision loss.
Green Fluorescent Protein notes.ppt
GFP has several advantages over other reporter genes for detecting gene expression:
1. GFP allows direct observation of gene expression over time in living cells, while other reporters require killing cells to measure expression.
2. GFP fluorescence can be measured at the single cell level by flow cytometry, providing quantitative data on variation in expression between individual cells.
3. Kinetic analysis of gene expression over time is possible by monitoring GFP fluorescence, which is not feasible with other reporters.
4. GFP fusions can be used to study protein localization and trafficking in living cells in real-time.
Transcription.ppt
Transcription is the process by which RNA polymerase converts DNA into RNA. It involves 3 stages: initiation, elongation, and termination. Initiation begins at promoter sequences on DNA and requires a sigma factor. During elongation, RNA polymerase proofreads as it synthesizes RNA to ensure accuracy. Termination occurs when the polymerase reaches termination sequences and releases the RNA transcript.
Bioenergetics.ppt
Catabolism breaks down molecules to release energy, while anabolism uses energy to build molecules. The three macronutrients that yield energy are carbohydrates, fats, and proteins. Carbohydrates have a limited storage capacity, fats have an unlimited storage capacity, and proteins are used for building with no storage capacity. Metabolic pathways are integrated as glucose, fatty acids, and amino acids are broken down into the common intermediate acetyl-CoA, then further oxidized, with degradation and synthesis usually compartmentalized. Glycolysis and the TCA cycle generate ATP through oxidation reactions regulated by key enzymes.
Green Fluorescent Protein.ppt
Green fluorescent protein (GFP) is a protein isolated from jellyfish that emits green light when exposed to blue light. It consists of 238 amino acid residues arranged in a beta-barrel structure with an alpha-helix running through the center. The fluorophore of GFP is formed by three amino acids in the central alpha-helix. GFP has become an important research tool, used as a marker for gene expression, protein localization and interactions by fusing it to genes or proteins of interest. It allows non-invasive visualization of biological processes using fluorescence microscopy.
Muscle Contraction and Movement_Enzymes.ppt
1. The document describes the molecular characteristics and mechanisms of muscle contraction. It discusses the structure and interactions of the contractile filaments actin and myosin.
2. The sliding filament theory is described as the mechanism where myosin heads bind to actin and hydrolyze ATP to slide along actin, causing muscle shortening.
3. The process of muscle contraction involves motor neuron stimulation, calcium ion release, actin-myosin cross-bridge formation, and sarcomere shortening through the sliding of actin and myosin filaments.
Prions ppt SBC 450.ppt
Prions are infectious protein particles that cause neurodegenerative diseases like Creutzfeldt-Jakob disease in humans and bovine spongiform encephalopathy (mad cow disease) in cattle. Prions are composed solely of protein and contain no nucleic acids. They propagate by converting the normal cellular prion protein (PrPC) into an abnormal disease-causing form (PrPSc). The normal prion protein is involved in the replication of the mutated form. Prion diseases are difficult to treat as prions are highly resistant to heat and chemical inactivation.
Tenancy agreement
This document appears to be a rental agreement between Mellick Solutions (the landlord's agent) and an unnamed tenant. The key details are:
1. The tenant agrees to rent part of a commercial/residential building identified as [blank] for an agreed monthly rent of [blank] payable by the 5th of each month.
2. The tenant pays an initial deposit equal to one month's rent.
3. The tenant agrees to only use the premises for [blank] purposes and not assign or sublet without permission. The tenant also agrees to pay utility bills and return the premises in good condition.
4. The landlord agrees to pay rates and head rents and allow the tenant peaceful
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学校原版美国波士顿大学毕业证学历学位证书原版一模一样
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3:国家专业人才认证中心颁发入库证书
4:这个认证书并且可以归档倒地方
5:凡事获得留信网入网的信息将会逐步更新到个人身份内，将在公安局网内查询个人身份证信息后，同步读取人才网入库信息
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SPR and Avidin-Biotin interaction notes.pptx
- 1. Surface plasmon resonance (SPR)-based biosensors • Surface plasmon resonance (SPR)-based biosensors are very powerful tools for the study of Label-free biomolecular interaction, chemical detection and immunoassays. • https://www.youtube.com/watch?v=sM-VI3alvAI • A biosensor is an analytical device for the analysis of biomaterial samples to gain an understanding of their bio-composition, structure and function by converting a biological response into an electrical signal It should basically consist of a transducer (electrochemical , piezoelectric, or optical) and a recognition element which captures a specific analyte. (check handout on electrochemical methods).
- 2. • Various optical sensing methods have been exploited in biosensors including chemiluminescence, fluorescence, light absorption and scattering, reflectance, and Surface plasmon resonance , which can be classified into two categories: label-based (the two former) and label- free (the others).
- 3. SPR biosensing principle: the analyte of interest is detected by measuring the change of the refractive index in the target solution.
- 5. • Avidin is a highly stable, tetrameric glycoprotein (molecular weight 66–69 kDa) that contains terminal N-acetyl glucosamine and mannose moieties. Each of the four subunits contains 128 amino acids and binds to biotin with high specificity and affinity • Avidin is originally derived from the eggs of aves, reptiles and amphibians. • Avidin-biotin interaction is considered one of the most specific and stable non-covalent interactions, which is about 103 to 106 times higher than an antigen-antibody interaction. • the strong interaction between avidin and biotin may pose a limitation in releasing the tagged biomolecules from the biotin or avidin
- 6. • Several genetically and chemically engineered avidin and its analogues have been studied to enhance knowledge about the functional and structural characteristics of avidins, which may lead to more successful applications. • The biggest advantage of this system is its high affinity interaction, which is robust and stable against manipulation, proteolytic enzymes, temperature, pH, harsh organic reagents, and other denaturing reagents. • Therefore, the avidin-biotin interaction serves as a great tool in the biomedical and nanotechnological applications. • On the other hand, biotin-based conjugates are easy to synthesize and have less impact on the activity of the biomolecules.
- 7. • Compared to other covalent and non-covalent interactions, the avidin-biotin system provides enormous advantages such as amplification of weak signals, efficient operation, highly stability and enables the use of highly diluted primary antibodies. • Therefore, avidin has been a very versatile modality in the field of biotechnology, especially biochemical assays and affinity purification, over four decades.
- 8. Biotin and analogues • Biotin is a vitamin also known as vitamin H, vitamin B7 or co-enzyme R. Biotin is composed of a tetrahydrothiophene ring fused to a tetrahydroimidizalone (ureido) ring. It plays a key role in cell signaling and acts as a cellular growth promoter. Biotin receptor (sodium-dependent multivitamin transporter and high-affinity biotin transporter) is widely expressed in nearly all living cells. • Moreover, its expression in dividing cancer cells is higher than in normal cells, making biotin a potential targeting moiety for cancer therapeutics. • Extensive effort has therefore been made to develop biotin-based platforms for tumor targeting and diagnosis. •
- 9. • The functional groups of biotin have been chemically modified to synthesize biotin analogues, such as iminobiotin, ethylbiotin, desthiobiotin, biotin-carbamate, and biotin-carbonate for various applications
- 10. Avidin-Biotin: Nature’s superglue • Exhibit highest known affinity in nature between a ligand and a protein- Ka=1015M-1 • Arises from hydrophobic interactions between biotin and aromatic amino acids arranged inside the avidin biding pockets • Avidin-biotin bonds are very difficult to break- pH 2-13, 9M Urea • Strongest non-covalent bond in nature • The avidin-biotin is highly specific and strong binding, the avidin-biotin system is widely used in a variety of biochemical applications Affinity chromatography Binding assays
- 11. Immobilizing a protein onto a biosensor using avidin-biotin bridges
- 12. Uses of avidin-biotin system • Applied to every type of biosensor and nanoparticle surface Optical Acoustic Electrochemical Nanoparticles • Detection of BIOTIN in an analyte solution using a sensor immobilized with AVIDIN is a standard test to simulate detection of PROTEIN-DRUG interactions High MW immobilized ligands Low MW analyte