Enzyme-Linked Immunosorbent Assay(ELISA)- 3rd BSc.pptx
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This document describes the Enzyme-Linked ImmunoSorbent Assay (ELISA) technique. ELISA uses an enzyme-substrate reaction to detect the presence of antigens or antibodies in specimens. There are four main types of ELISA - direct, indirect, sandwich, and competitive - which differ based on whether antigens or antibodies are being detected and the order that reaction components are added. ELISA is a sensitive and specific technique that allows for high-throughput testing using microtiter plates, but requires specialized equipment. It has various applications for detecting viruses, bacteria, and other pathogens.
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ELISA.pptx
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Presentation elisa ppt
ELISA is an immunological technique used to detect the presence of antigens or antibodies in samples. It works by immobilizing an antigen or antibody on a plate and detecting it using an enzyme-linked antibody or antigen. This allows quantification of the analyte being tested for. ELISA has advantages like being sensitive, specific, easy to perform, and having a long shelf life for reagents. It is commonly used to detect proteins, hormones, drugs, tumor markers, antibodies, and antigens in clinical and research applications.
ELISA TECHNIQUE
This document discusses enzyme-linked immunosorbent assay (ELISA), a sensitive technique used to detect antigens or antibodies in biological samples. It describes different types of ELISA including direct, indirect, sandwich, and competitive ELISA. The key steps of ELISA are reviewed including coating wells with antigens or antibodies, adding samples and enzyme-labeled antibodies, and detecting color change using enzyme substrates and chromogens. Advantages of ELISA are its sensitivity, ability to test many samples simultaneously, and common use in screening tests. Disadvantages include the need for specialized equipment and higher cost than rapid tests for individual samples.
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The document describes several types of enzyme-linked immunosorbent assays (ELISA) including direct ELISA, indirect ELISA, sandwich ELISA, and competitive ELISA. ELISA uses an enzyme-linked antibody or antigen to detect or quantify a target molecule in samples. The rate of color change when a substrate is added is proportional to the amount of target molecule present. Atomic absorption spectroscopy also quantifies metals by generating atomic vapors and measuring their light absorption at specific wavelengths.
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ELISA (Enzyme-Linked Immunosorbent Assay) is a widely used technique for detecting antigens or antibodies. It involves coating a microtiter plate with an antigen or antibody and detecting its binding with an enzyme-linked secondary antibody. There are several types of ELISA including direct, indirect, sandwich, and competitive ELISA, which differ in their detection methods and targets. ELISA has many applications, such as detecting antibodies for diseases like tuberculosis, hepatitis B, and HIV, as well as detecting food allergens and bacterial toxins.
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This document provides an overview of ELISA (enzyme-linked immunosorbent assay), including its principle, evolution, types, basic steps, and applications. ELISA is an immunoassay technique that detects and quantifies substances like proteins, antibodies, and hormones. It relies on antibodies and antigens binding specifically together, and uses enzyme labeling to identify these bindings colorimetrically. The document discusses various ELISA techniques like direct ELISA, indirect ELISA, and sandwich ELISA.
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ELISA is an immunological technique used to detect the presence of antigens or antibodies in samples. It works by immobilizing an antigen or antibody on a plate and detecting it using an enzyme-linked antibody or antigen. This allows quantification of the analyte being tested for. ELISA has advantages like being sensitive, specific, easy to perform, and having a long shelf life for reagents. It is commonly used to detect proteins, hormones, drugs, tumor markers, antibodies, and antigens in clinical and research applications.
ELISA TECHNIQUE
This document discusses enzyme-linked immunosorbent assay (ELISA), a sensitive technique used to detect antigens or antibodies in biological samples. It describes different types of ELISA including direct, indirect, sandwich, and competitive ELISA. The key steps of ELISA are reviewed including coating wells with antigens or antibodies, adding samples and enzyme-labeled antibodies, and detecting color change using enzyme substrates and chromogens. Advantages of ELISA are its sensitivity, ability to test many samples simultaneously, and common use in screening tests. Disadvantages include the need for specialized equipment and higher cost than rapid tests for individual samples.
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The document describes several types of enzyme-linked immunosorbent assays (ELISA) including direct ELISA, indirect ELISA, sandwich ELISA, and competitive ELISA. ELISA uses an enzyme-linked antibody or antigen to detect or quantify a target molecule in samples. The rate of color change when a substrate is added is proportional to the amount of target molecule present. Atomic absorption spectroscopy also quantifies metals by generating atomic vapors and measuring their light absorption at specific wavelengths.
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ELISA (Enzyme-Linked Immunosorbent Assay) is a widely used technique for detecting antigens or antibodies. It involves coating a microtiter plate with an antigen or antibody and detecting its binding with an enzyme-linked secondary antibody. There are several types of ELISA including direct, indirect, sandwich, and competitive ELISA, which differ in their detection methods and targets. ELISA has many applications, such as detecting antibodies for diseases like tuberculosis, hepatitis B, and HIV, as well as detecting food allergens and bacterial toxins.
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The document describes the ELISA (enzyme-linked immunosorbent assay) technique. ELISA uses antibodies and enzymes to detect the presence of antigens or antibodies in a liquid sample. It involves attaching an antigen or antibody to a solid surface and using a labeled antibody or antigen to detect binding. The enzyme label converts a colorless substrate to a colored product, indicating the presence of the antigen or antibody being tested for. There are several types of ELISA including direct, indirect, sandwich, and competitive ELISA. ELISA has various applications including screening blood donations, measuring hormone levels, and detecting infections.
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The ELISA (enzyme-linked immunosorbent assay) is a popular biochemistry assay that uses antibodies and color change to identify a substance. It involves using an enzyme-linked antibody to detect antigen-antibody binding, where the enzyme converts a colorless substrate into a colored product. There are several types of ELISA including indirect, direct, sandwich, and competitive ELISA. ELISA has various applications such as screening blood donations, measuring hormone levels, and detecting infections and allergens.
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ELISA, or enzyme-linked immunosorbent assay, are immunological procedures in which the Ag- Ab reaction is monitored by enzyme measurements
ELISA (enzyme-linked immuno sorbent assay) by Pranzly.ppt
INTRODUCTION
The term ELISA was first used by Engvall & Perlma in 1971.
high sensitivity
useful & powerful method in estimating ng/mL to pg/mL ordered materials in the solution.
Similar To RIA, Except No Radio-labelling.
Alkaline phosphatase, horseradish peroxidase and beta-galactosidase are the enzymes used in the EIA tests.
PRINCIPLE
MATERIAL REQUIRED
REAGENTS
TYPES
NON-COMPETITIVE ELISA
DIRECT ELISA
INDIRECT ELISA
SANDWICH ELISA
COMPETITIVE ELISA
ELISA RESULT
QUALITATIVE
QUANTITATIVE
SEMI-QUANTITATIVE
PRECAUTIONS
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The enzyme-linked immunosorbent assay (ELISA) is a common laboratory technique used to measure the concentration of an analyte like antibodies or antigens in solution. It works by using the principle of antigen-antibody binding and an enzyme-linked secondary antibody or antigen to detect the presence of an analyte. There are several types of ELISA including direct, indirect, sandwich, and competitive based on the procedure and detection method used. ELISA is a sensitive and specific technique with advantages like using inexpensive and stable reagents, but it also has limitations like possible false positives and negatives.
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This document provides an overview of ELISA (enzyme-linked immunosorbent assay), a commonly used laboratory test to detect antibodies in blood. It discusses the basic principles of antigen-antibody interactions, the requirements and basic procedure for an ELISA test, and the four main types: direct ELISA, indirect ELISA, sandwich ELISA, and competition ELISA. ELISA testing is used to diagnose various infectious diseases and conditions by detecting antibodies produced in response to antigens. The procedure involves immobilizing antigens, adding samples and detection antibodies, conjugating enzymes to antibodies, and measuring color change from substrates to quantify the target protein.
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The document provides guidelines for performing ELISA (Enzyme-Linked Immunosorbent Assay), including:
1. ELISA is a popular immunological technique that uses antibodies and enzymes to detect antigens or antibodies. It is very sensitive and can be used both qualitatively and quantitatively.
2. The basic steps of ELISA involve coating a plate with an antigen, adding a primary antibody, adding a secondary antibody linked to an enzyme, adding a substrate that reacts with the enzyme to produce a colored product.
3. Factors that can cause troubleshooting issues include contamination, improper washing, expired reagents, incorrect procedures, temperature issues, poor pipetting technique, and more. Proper controls and validation
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The ELISA (enzyme-linked immunosorbent assay) is a test that uses antibodies and color change to identify a substance. It involves using an enzyme to detect antigen-antibody binding, which converts a colorless substrate into a colored product. There are several types of ELISA including indirect, direct, sandwich, and competitive ELISA. ELISA can provide quantitative or qualitative results and has applications like screening donated blood and measuring hormone levels.
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The document provides an overview of enzyme-linked immunosorbent assay (ELISA), including its history, principles, requirements, types (direct, indirect, sandwich, competitive), applications (ELISPOT), equipment (washer, reader), and generations used for HIV detection. ELISA is a sensitive technique for detecting antigen-antibody interactions using an enzyme-substrate system for detection. It was developed in 1971 and involves multiple steps with different reagents to separate bound and unbound fractions.
Enzyme Linked Immunosorbent Assay
ELISA (enzyme-linked immunosorbent assay) is an analytical biochemistry technique used to detect the presence of an antibody or antigen in a sample using antibodies specific to the target antigen or antibody. There are four main types of ELISA: direct, indirect, sandwich, and competitive. ELISA uses an enzyme-linked antibody or antigen to detect the target molecule and a chromogenic substrate to produce a detectable color change that corresponds to the amount of target molecule present. ELISA has many applications in clinical diagnostics, pharmaceutical analysis, and food safety due to its high sensitivity and specificity.
ELISA
ELISA (enzyme-linked immunosorbent assay) is a sensitive analytical method for detecting low amounts of substances like proteins, peptides, hormones, and infectious agents. It uses antibodies and enzyme-labeled detection molecules to detect the presence of an antigen or antibody in a sample. There are two main types - sandwich ELISA, which is most sensitive and requires the analyte to have at least two binding sites, and competitive ELISA, which is useful for monovalent analytes. ELISA has advantages like specificity, sensitivity, and ease of use, though limitations include potential for false positives and negatives. It has many applications like detecting hormones, proteins, infectious agents, and more.
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Elisa
ELISA, or enzyme-linked immunosorbent assay, is a quantitative immunological technique used to detect the presence of antigens or antibodies in a sample. In ELISA, an enzyme is used to detect the binding of an antigen to its antibody. The enzyme then converts a colorless substrate into a colored product, indicating the presence of the antigen-antibody binding. ELISA can be used for both qualitative detection and quantitative measurement of antigens or antibodies. Common enzymes used in ELISA include horseradish peroxidase, alkaline phosphatase, and β-galactosidase.
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The document discusses the ELISA (enzyme-linked immunosorbent assay) technique. ELISA is used to detect the presence and quantity of antigens or antibodies in samples. It works by capturing the target antigen or antibody using specific antibodies attached to a microwell plate. An enzyme-linked antibody is then used to detect the captured target, and a color change reaction indicates a positive result. Common applications include detecting hormones, viruses, bacteria, and antibodies in blood samples. There are two main types: indirect ELISA detects antibodies, while sandwich ELISA detects antigens.
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Radioimmunoassay (RIA) is an assay technique that uses radioisotopes rather than enzymes as labels to detect and quantify antigens or antibodies. In RIA, a radioisotope is attached to an antigen and combined with an antibody. A sample containing the target antigen is then added and competes with the radioactive antigen for binding sites on the antibody. The amount of unbound radioactive antigen is then measured, with higher levels of the target antigen resulting in lower radioactivity, allowing very small quantities to be detected. RIA is highly specific and can be used to diagnose diseases or measure hormones.
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The ELISA (enzyme-linked immunosorbent assay) is a popular biochemistry assay that uses antibodies and color change to identify a substance. It involves using an enzyme-linked antibody to detect antigen-antibody binding, where the enzyme converts a colorless substrate into a colored product. There are several types of ELISA including indirect, direct, sandwich, and competitive ELISA. ELISA has various applications such as screening blood donations, measuring hormone levels, and detecting infections and allergens.
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The document summarizes the enzyme-linked immunosorbent assay (ELISA) technique. It describes the basic process which involves coating a solid surface like a microwell plate with an antigen or antibody, adding an enzyme-linked antibody or antigen to detect binding, washing away unbound reagents, and measuring the enzyme activity using a chromogenic substrate which produces a detectable color change. It discusses different types of ELISA based on detection methods or the antigen-antibody binding procedure. It also lists materials commonly used and provides some advantages and limitations of the ELISA technique.
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ELISA, or enzyme-linked immunosorbent assay, are immunological procedures in which the Ag- Ab reaction is monitored by enzyme measurements
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INTRODUCTION
The term ELISA was first used by Engvall & Perlma in 1971.
high sensitivity
useful & powerful method in estimating ng/mL to pg/mL ordered materials in the solution.
Similar To RIA, Except No Radio-labelling.
Alkaline phosphatase, horseradish peroxidase and beta-galactosidase are the enzymes used in the EIA tests.
PRINCIPLE
MATERIAL REQUIRED
REAGENTS
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The enzyme-linked immunosorbent assay (ELISA) is a common laboratory technique used to measure the concentration of an analyte like antibodies or antigens in solution. It works by using the principle of antigen-antibody binding and an enzyme-linked secondary antibody or antigen to detect the presence of an analyte. There are several types of ELISA including direct, indirect, sandwich, and competitive based on the procedure and detection method used. ELISA is a sensitive and specific technique with advantages like using inexpensive and stable reagents, but it also has limitations like possible false positives and negatives.
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This document provides an overview of ELISA (enzyme-linked immunosorbent assay), a commonly used laboratory test to detect antibodies in blood. It discusses the basic principles of antigen-antibody interactions, the requirements and basic procedure for an ELISA test, and the four main types: direct ELISA, indirect ELISA, sandwich ELISA, and competition ELISA. ELISA testing is used to diagnose various infectious diseases and conditions by detecting antibodies produced in response to antigens. The procedure involves immobilizing antigens, adding samples and detection antibodies, conjugating enzymes to antibodies, and measuring color change from substrates to quantify the target protein.
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1. ELISA is a popular immunological technique that uses antibodies and enzymes to detect antigens or antibodies. It is very sensitive and can be used both qualitatively and quantitatively.
2. The basic steps of ELISA involve coating a plate with an antigen, adding a primary antibody, adding a secondary antibody linked to an enzyme, adding a substrate that reacts with the enzyme to produce a colored product.
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ELISA (enzyme-linked immunosorbent assay) is an analytical biochemistry technique used to detect the presence of an antibody or antigen in a sample using antibodies specific to the target antigen or antibody. There are four main types of ELISA: direct, indirect, sandwich, and competitive. ELISA uses an enzyme-linked antibody or antigen to detect the target molecule and a chromogenic substrate to produce a detectable color change that corresponds to the amount of target molecule present. ELISA has many applications in clinical diagnostics, pharmaceutical analysis, and food safety due to its high sensitivity and specificity.
ELISA
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Elisa
ELISA, or enzyme-linked immunosorbent assay, is a quantitative immunological technique used to detect the presence of antigens or antibodies in a sample. In ELISA, an enzyme is used to detect the binding of an antigen to its antibody. The enzyme then converts a colorless substrate into a colored product, indicating the presence of the antigen-antibody binding. ELISA can be used for both qualitative detection and quantitative measurement of antigens or antibodies. Common enzymes used in ELISA include horseradish peroxidase, alkaline phosphatase, and β-galactosidase.
ELISA
The document discusses the ELISA (enzyme-linked immunosorbent assay) technique. ELISA is used to detect the presence and quantity of antigens or antibodies in samples. It works by capturing the target antigen or antibody using specific antibodies attached to a microwell plate. An enzyme-linked antibody is then used to detect the captured target, and a color change reaction indicates a positive result. Common applications include detecting hormones, viruses, bacteria, and antibodies in blood samples. There are two main types: indirect ELISA detects antibodies, while sandwich ELISA detects antigens.
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- Lipid digestion requires emulsification by bile and lipolysis by pancreatic lipase to break triglycerides into fatty acids and monoglycerides absorbed via micelles. Conditions like lactose intolerance and malabsorption syndrome can
LFT.pptx
This document discusses liver function tests. It classifies tests based on laboratory findings and clinical aspects. Key tests include bilirubin to measure excretory function, albumin and prothrombin time to measure synthetic function, and ALT/AST to measure hepatocellular injury. Alkaline phosphatase and GGT indicate obstructive liver disease or cholestasis. Normal ranges are provided for important tests like bilirubin, ALT, AST, alkaline phosphatase. Methods for measuring bilirubin and calculating A:G ratio are also outlined.
Progressive systemic sclerosis
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Breast cancer: Post menopausal endocrine therapy
Breast cancer in postmenopausal women with hormone receptor-positive (HR+) status is a common and complex condition that necessitates a multifaceted approach to management. HR+ breast cancer means that the cancer cells grow in response to hormones such as estrogen and progesterone. This subtype is prevalent among postmenopausal women and typically exhibits a more indolent course compared to other forms of breast cancer, which allows for a variety of treatment options.
Diagnosis and Staging
The diagnosis of HR+ breast cancer begins with clinical evaluation, imaging, and biopsy. Imaging modalities such as mammography, ultrasound, and MRI help in assessing the extent of the disease. Histopathological examination and immunohistochemical staining of the biopsy sample confirm the diagnosis and hormone receptor status by identifying the presence of estrogen receptors (ER) and progesterone receptors (PR) on the tumor cells.
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Endocrine Therapy
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Selective Estrogen Receptor Modulators (SERMs): Tamoxifen is a SERM that binds to estrogen receptors, blocking estrogen from stimulating breast cancer cells. It is effective but may have side effects such as increased risk of endometrial cancer and thromboembolic events.
Aromatase Inhibitors (AIs): These drugs, including anastrozole, letrozole, and exemestane, lower estrogen levels by inhibiting the aromatase enzyme, which converts androgens to estrogen in peripheral tissues. AIs are generally preferred in postmenopausal women due to their efficacy and safety profile compared to tamoxifen.
Selective Estrogen Receptor Downregulators (SERDs): Fulvestrant is a SERD that degrades estrogen receptors and is used in cases where resistance to other endocrine therapies develops.
Combination Therapies
Combining endocrine therapy with other treatments enhances efficacy. Examples include:
Endocrine Therapy with CDK4/6 Inhibitors: Palbociclib, ribociclib, and abemaciclib are CDK4/6 inhibitors that, when combined with endocrine therapy, significantly improve progression-free survival in advanced HR+ breast cancer.
Endocrine Therapy with mTOR Inhibitors: Everolimus, an mTOR inhibitor, can be added to endocrine therapy for patients who have developed resistance to aromatase inhibitors.
Chemotherapy
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Cell Therapy Expansion and Challenges in Autoimmune Disease
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Widespread adoption of cell therapies will not only require strong efficacy and safety data, but also adapted pricing and access strategies. At oncology-based price points, CAR-Ts are unlikely to achieve broad market access in autoimmune disorders, with eligible patient populations that are potentially orders of magnitude greater than the number of currently addressable cancer patients. Developers have made strides towards reducing cell therapy COGS while improving manufacturing efficiency, but payors will inevitably restrict access until more sustainable pricing is achieved.
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- 2. •Enzyme immunoassay (EIA) is a term used to describe all test that detect either Ag/Ab in the specimens by using enzyme-substrate system . • The enzyme reacts with the substrate, which in turn activates chromogen to produce a color. • The colour change is detected by spectrophotometry
- 3. •Enzyme: used to label one of the component ie. Ag/Ab. •Substrate-chromogen is added at the final step.
- 4. ENZYME SUBSTRATE CHROMOGEN Horseradish peroxide Hydrogen peroxide Tetramethyl benzidine • Performed in MICROTITRE PLATE
- 6. Types of ELISA Direct ELISA Indirect ELISA Sandwich ELISA Competitive ELISA
- 7. Direct ELISA •Detection of Ag in test serum. •Primary Ab is labeled with enzyme.
- 8. (In serum)
- 9. •The color change is detected by spectrophotometry
- 10. Indirect ELISA •Used for Detection of Ab •Secondary Ab is labeled with enzyme
- 11. (In serum)
- 12. •The colour change is detected by spectrophotometry
- 13. Sandwich ELISA • Detection of Ag in test serum. • The Ag gets sandwiched between capture Ab and detector Ab
- 14. STEPS: Well coated with capture Ab +Ag(test serum)+ Primary Ab-enzyme + substrate chromogen color changes • The color change is detected by spectrophotometry
- 16. Competitive ELISA The Ag in test serum compete with another Ag of same type, coated on well to bind to the primary Ab.
- 17. STEPS: oPrimary Ab is incubated in a solution with serum sample with Ag. oThe Ag-Ab mixture is added to microtitre well precoated with same type of Ag. oThe free Ab bind to the Ag coated in the well. oEnzyme conjugated 2nd Ab is added •substrate- chromogen is added, The color change is detected by spectrophotometry
- 19. Advantages of ELISA Large number of samples can be tested. More sensitive. More specific
- 20. Disadvantages •Expensive equipments are required
- 21. APPLICATIONS • Ag detection : HepB –HBsAg, HBeAg Dengue –NS1 Ag • Ab detection : HIV, EBV, HepB, HepC