This document discusses various immune assays and their applications, focusing on enzyme-linked immunosorbent assays (ELISA). It describes the basic principles and components of ELISA, including indirect, sandwich, and competitive ELISA techniques. Applications of ELISA include screening blood donations, detecting infections, measuring hormone and toxin levels, and detecting illicit drugs. Neutralization and precipitation are also summarized as methods to identify toxins/antitoxins and detect antigen-antibody reactions.
ELISA, Principles of ELISA, Types of ELISA- Direct ELISA
Indirect ELISA, Sandwich ELISA, Competitive ELISA, and other Types i.e. ELISPOT (enzyme-linked immunospot assay) and In-cell ELISA, Advantages and disadvantages of ELISA detection methods, Different types of microplates for ELISA, Detection strategies for ELISA
ELISA, Principles of ELISA, Types of ELISA- Direct ELISA
Indirect ELISA, Sandwich ELISA, Competitive ELISA, and other Types i.e. ELISPOT (enzyme-linked immunospot assay) and In-cell ELISA, Advantages and disadvantages of ELISA detection methods, Different types of microplates for ELISA, Detection strategies for ELISA
Enzyme-Linked ImmunoSorbent Assay, or ELISA, is a biochemical technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample. The ELISA has been used as a diagnostic tool in medicine and plant pathology, as well as a quality control check in various industries. In simple terms, in ELISA an unknown amount of antigen is affixed to a surface, and then a specific antibody is washed over the surface so that it can bind the antigen. This antibody is linked to an enzyme, and in the final step a substance is added that the enzyme can convert to some detectable signal.
Immunodiffusion -Different Types,Principle,procedureand application. it is a diagnostic technique for the detection or measurements of antibodies and antigens by their precipitation which involves diffusion through a substances such as agar or gel agarose .common types -oudin procedure,oakley fulthorpe procedure ,mancini technique ,ouchterlony double immuno diffusion
A path breaking technology which has made it possible for us to detect HIV. ELISA is an immunological assay nowadays even used to detect food proteins & is the science behind pregnancy color test. This presentation unlocks the working of this assay.
Enzyme immunoassays (EIAs), also known as enzyme-linked immunosorbent assays (ELISAs), combine antibody binding with enzymatic detection to quantify molecules of interest.
Test for detection of plant virus by ELISA test.pdfLOKESH R
This presentation provides an overview of the Enzyme-Linked Immunosorbent Assay (ELISA) test for the detection of plant viruses. The ELISA test is a highly sensitive and specific method for detecting viral antigens in plant tissues, seeds, and other plant materials.
The presentation covers the principles of ELISA test, including the different types of ELISA tests, such as direct, indirect, sandwich, and competitive ELISA. The steps involved in the ELISA test, including sample preparation, antibody labeling, incubation, and detection, are discussed in detail.
The advantages and limitations of the ELISA test for plant virus detection are also highlighted. The presentation includes a comparison of the ELISA test with other diagnostic methods for plant viruses, such as polymerase chain reaction (PCR) and serological assays.
Finally, the presentation provides examples of the applications of the ELISA test for plant virus detection, including its use in crop protection, disease management, and quarantine measures. The presentation concludes with a summary of the key points discussed and recommendations for the use of the ELISA test in plant virus detection.
Enzyme-Linked ImmunoSorbent Assay, or ELISA, is a biochemical technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample. The ELISA has been used as a diagnostic tool in medicine and plant pathology, as well as a quality control check in various industries. In simple terms, in ELISA an unknown amount of antigen is affixed to a surface, and then a specific antibody is washed over the surface so that it can bind the antigen. This antibody is linked to an enzyme, and in the final step a substance is added that the enzyme can convert to some detectable signal.
Immunodiffusion -Different Types,Principle,procedureand application. it is a diagnostic technique for the detection or measurements of antibodies and antigens by their precipitation which involves diffusion through a substances such as agar or gel agarose .common types -oudin procedure,oakley fulthorpe procedure ,mancini technique ,ouchterlony double immuno diffusion
A path breaking technology which has made it possible for us to detect HIV. ELISA is an immunological assay nowadays even used to detect food proteins & is the science behind pregnancy color test. This presentation unlocks the working of this assay.
Enzyme immunoassays (EIAs), also known as enzyme-linked immunosorbent assays (ELISAs), combine antibody binding with enzymatic detection to quantify molecules of interest.
Test for detection of plant virus by ELISA test.pdfLOKESH R
This presentation provides an overview of the Enzyme-Linked Immunosorbent Assay (ELISA) test for the detection of plant viruses. The ELISA test is a highly sensitive and specific method for detecting viral antigens in plant tissues, seeds, and other plant materials.
The presentation covers the principles of ELISA test, including the different types of ELISA tests, such as direct, indirect, sandwich, and competitive ELISA. The steps involved in the ELISA test, including sample preparation, antibody labeling, incubation, and detection, are discussed in detail.
The advantages and limitations of the ELISA test for plant virus detection are also highlighted. The presentation includes a comparison of the ELISA test with other diagnostic methods for plant viruses, such as polymerase chain reaction (PCR) and serological assays.
Finally, the presentation provides examples of the applications of the ELISA test for plant virus detection, including its use in crop protection, disease management, and quarantine measures. The presentation concludes with a summary of the key points discussed and recommendations for the use of the ELISA test in plant virus detection.
This presentation explains about the principle and procedure involved in elisa method of immunoassay, development o f elisa , application advantages and disadvantages of elisa
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
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Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Francesca Gottschalk - How can education support child empowerment.pptxEduSkills OECD
Francesca Gottschalk from the OECD’s Centre for Educational Research and Innovation presents at the Ask an Expert Webinar: How can education support child empowerment?
Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
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3. IMMUNE ASSAYS Immunoassays are a group of sensitive analytical tests that utilize very specific antibody/antigen complexes to produce a signal that can be measured and related to the concentration of a compound in solution. TYPES Radioimmunoassays(RIAs) Fluorescent Polarized Immunoassay Enzyme Multiplied Immunoassay (EMIT) Enzyme linked immunosorbant assay (ELISA) 11 May 2002 3
5. INTRODUCTION TO ELISA ELISA, or enzyme-linked immunosorbent assay, is an immunoassay technique involving the reaction of antigen and antibody in vitro. ELISA is a sensitive and specific assay for the detection and quantitation of antigens or antibodies. ELISA tests are usually performed in microwell plates. It is a sensitive immunoassay that uses an enzyme linked to an antibody or antigen as a marker for the detection of a specific protein. 11 May 2002 5
6. HISTORY It is first described by Peter Perlmannand Eva Engval , and Anton SchuursandBauke van Weemen. The technique was subsequently developed by Voller and col. using microplates, that permitted the development of highly sensitive and accurate kits. 11 May 2002 6
7. COMPONENTS OF AN ELISA Antibody: IgG fraction of serum Enzyme:Peroxidase from horseradish,Alkalinephosphatase from E. coli,β-galactosidase from E coli.,Glucoseoxidase. Substrate: TMB (3,3',5,5', tetramethylbenzidine). 11 May 2002 7
8. PRINCIPLE OF ELISA The sample with an unknown amount of antigen is immobilized on a solid support. The detection antibody is added ,forming a complex with antigen. The detection antibody can be covalently linked to an enzyme, or can itself be detected by a secondary antibody which is linked to an enzyme. Between each step the plate washed with a mild detergent solution. After the final wash step, adding an enzymatic substrate to produce a visible signal, which indicates the quantity of antigen in the sample. 11 May 2002 8
9. 8. Observe colour development 7. Add substrate for enzyme 1. Add antigen 2. Wash with PBST 6. Wash with PBST 4. Wash with PBST 3. Add primary antibody 5. Add secondary antibody 11 May 2002 9
10. ELISA Step 1 inactivated HIV antigens Step 2 serum antibodies Step 3 Anti-human Ig coupled to enzyme Step 4 Chromogen or substrate Step 5 ( Measurement )
11. CRITERIA FOR CHOICE OF A MARKER ENZYME Should be easily coupled to ligands & the labelled complex must be stable. The reactivity should be retained after linking of the enzyme to the antigen/antibody. The chosen enzymes should not be normally present in the patient samples. 11 May 2002 11
12. TYPES OF ELISA INDIRECT ELISA SANDWICH ELISA COMPETETIVE ELISA 11 May 2002 12
13. INDIRECT ELISA The indirect ELISA utilizes an unlabeled primary antibody in conjunction with a labeled secondary antibody. Since first the antigen is coated and specific antibody is added which forms complex.Then add enzyme-conjugated secondry antibody and add substrate and measure colour. 11 May 2002 13
14. ADVANTAGES AND DISDVANTAGES Advantages of indirect detection Wide variety of labeled secondary antibodies are available commercially. Sensitivity is increased because each primary antibody contains several sites that can be bound by the labeled secondary antibody, allowing for signal amplification. Secondary-antibody conjugate avoids the expensive process of creating enzyme-linked antibodies for every antigen one might want to detect Disadvantages of indirect detection Cross-reactivity may occur with the secondary antibody, resulting in nonspecific signal. An extra incubation step is required in the procedure. 11 May 2002 14
15. SANDWICH ELISA Plate is coated with a antibody. Sample is added, and any antigen present binds to antibody. Detecting antibody is added, and binds to antigen. Enzyme-linked secondary antibody is added, and binds to detecting antibody. Substrate is added, and is converted by enzyme to detectable form. 11 May 2002 15
16. Competitive binding assay Unlabeled antibody is incubated in the presence of its antigen. These bound antibody/antigen complexes are then added to an antigen coated well. The plate is washed, so that unbound antibody is removed. (The more antigen in the sample, the less antibody will be able to bind to the antigen in the well, hence "competition.") The secondary antibody, specific to the primary antibody is added. This second antibody is coupled to the enzyme. A substrate is added and check the colour change. 11 May 2002 16
17. Advantages The major advantage of a competitive ELISA is the ability to use crude or impure samples and still selectively bind any antigen that may be present. 11 May 2002 17
19. APPLICATIONS Screening donated blood for evidence of viral contamination by HIV-1 and HIV-2 (presence of anti-HIV antibodies) hepatitis C (presence of antibodies) hepatitis B (testing for both antibodies and a viral antigen) Measuring hormone levels HCG (as a test for pregnancy) LH (determining the time of ovulation) TSH, T3 and T4 (for thyroid function) 11 May 2002 19
20. Detecting infections sexually-transmitted agents like HIV, syphilis and chlamydia hepatitis B and C Toxoplasmagondii Detecting allergens in food and house dust Measuring toxins in contaminated food Detecting illicit drugs, e.g., cocaine opiates 11 May 2002 20
31. NEUTRALIZATION It is a serological test used to identify used to identify toxins and antitoxins as well as viruses and viral antibodies. It involves antigen-antibody reaction. Laboratory animals are used as “indicator systems” in these tests. For example it is used to detect exotoxin of Clostridium botulinumin food. 11 May 2002 25
32.
33. In this test diphtheria toxin is injected intradermally.No skin reactions occurs if person has neutralizing antibodies. A local edema occurs if no neutralizing antibodies are present , indicating person is susceptible to diphtheria. 11 May 2002 26
35. PRECIPITATION The immunoprecipitation technique detects soluble antigens that react with antibodies called precipitins. The antibodies link the antigen to form a large antibody-antigen network or lattice that settles out of solution at the equivalence zone when it becomes sufficiently large. In fluids,antigen and antibody are layered over each other in a thin tube.The molecules then diffuse through the fluid untill they reach equivalence zone.A visible mass of particles is now formed at interface or at bottom. 11 May 2002 28
36.
37. Antigen and antibody solutions are placed in wells cut into agar in petridishes.The plates are incubated and precipitation lines form at zone of equivalence.
38. Used to detect fungal antigens of histoplasma,blastomyces,coccidioides.11 May 2002 29