This document provides an overview of enzyme-linked immunosorbent assay (ELISA), including its basic principles, types (sandwich, indirect, direct, competitive), applications (measuring hormones, detecting infections, allergens), and components (solid phase, adsorption, washing, antigen, antibody, enzyme conjugate, chromogen). ELISA is an important immunological method for detecting antigens and antibodies based on an enzyme-antibody or antigen-antibody reaction. It has advantages over radioimmunoassay such as higher sensitivity, safety, lower cost, and simpler instrumentation.
ELISA, principle and method by kk sahuKAUSHAL SAHU
What is ELISA.
Principle.
History.
Types of ELISA method.
1.Direct ELISA.
2.Indirect ELISA.
3.Sandwhich ELISA.
Conclusion.
References.
Antibodies (also known as immunoglobulins abbreviated Ig) are gamma globulin proteins that are found in blood and are used by the immune system to identify and neutralize foreign objects, such as bacteria and viruses.
Enzyme immunoassays (EIAs), also known as enzyme-linked immunosorbent assays (ELISAs), combine antibody binding with enzymatic detection to quantify molecules of interest.
ELISA, principle and method by kk sahuKAUSHAL SAHU
What is ELISA.
Principle.
History.
Types of ELISA method.
1.Direct ELISA.
2.Indirect ELISA.
3.Sandwhich ELISA.
Conclusion.
References.
Antibodies (also known as immunoglobulins abbreviated Ig) are gamma globulin proteins that are found in blood and are used by the immune system to identify and neutralize foreign objects, such as bacteria and viruses.
Enzyme immunoassays (EIAs), also known as enzyme-linked immunosorbent assays (ELISAs), combine antibody binding with enzymatic detection to quantify molecules of interest.
Hormones, Proteins, etc. present in blood in minute concentration can be assayed by the recent advanced technique of “Enzyme Immuno Assay” without involving any disadvantage. The basic reaction is the interaction between an antibody and an antigen.
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
ABSTRACT: The ELISA technique is a simple, sensitive, rapid, reliable, and versatile assay system for the quantitation of antigens and antibodies. Because of the extreme discriminating power of antibodies to recognize an almost infinite array of antigenic structures, the application of ELISA to analyte measurement is almost unlimited. ELISAs have been developed in many configurations depending on the particular application of the assay.
In solid-phase ELISA, one of the immunoreactants (antibody or antigen) is immobilized onto a solid support (microtiter plate) by adsorption, through non-covalent interactions. The immobilized antibody is then incubated with test solution containing the analyte of interest. Following a period of incubation and washing, the bound antigen is detected, by the addition of an enzyme-conjugated antibody that binds to the remaining antigenic sites on the antigen.
Although the technique is easy to perform and quite sensitive, there are certain problems to be solved before it becomes widely usable. In the present Memorandum the technical details are given and the advantages and shortcomings of the procedure are discussed. Present applications and future prospects are reviewed.
what is sandwich elisa, introduction to elisa, its type and main focus on sandwich elisa, , its process and advantages along with the disadvantages, its applications
Introduction, the principle of immunofluorescence, Technique, Fluorescent microscope and its components, Application and types of immunofluorescence, Direct and indirect immunofluorescence, FACS (Fluorescence-activated cell sorting), Uses and limitations of Immunofluorescence
Hormones, Proteins, etc. present in blood in minute concentration can be assayed by the recent advanced technique of “Enzyme Immuno Assay” without involving any disadvantage. The basic reaction is the interaction between an antibody and an antigen.
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
ABSTRACT: The ELISA technique is a simple, sensitive, rapid, reliable, and versatile assay system for the quantitation of antigens and antibodies. Because of the extreme discriminating power of antibodies to recognize an almost infinite array of antigenic structures, the application of ELISA to analyte measurement is almost unlimited. ELISAs have been developed in many configurations depending on the particular application of the assay.
In solid-phase ELISA, one of the immunoreactants (antibody or antigen) is immobilized onto a solid support (microtiter plate) by adsorption, through non-covalent interactions. The immobilized antibody is then incubated with test solution containing the analyte of interest. Following a period of incubation and washing, the bound antigen is detected, by the addition of an enzyme-conjugated antibody that binds to the remaining antigenic sites on the antigen.
Although the technique is easy to perform and quite sensitive, there are certain problems to be solved before it becomes widely usable. In the present Memorandum the technical details are given and the advantages and shortcomings of the procedure are discussed. Present applications and future prospects are reviewed.
what is sandwich elisa, introduction to elisa, its type and main focus on sandwich elisa, , its process and advantages along with the disadvantages, its applications
Introduction, the principle of immunofluorescence, Technique, Fluorescent microscope and its components, Application and types of immunofluorescence, Direct and indirect immunofluorescence, FACS (Fluorescence-activated cell sorting), Uses and limitations of Immunofluorescence
This presentation explains about the principle and procedure involved in elisa method of immunoassay, development o f elisa , application advantages and disadvantages of elisa
What is enzyme-linked immunosorbent assay?
A laboratory technique that uses antibodies linked to enzymes to detect and measure the amount of a substance in a solution, such as serum. The test is done using a solid surface to which the antibodies and other molecules stick.
ELISA (enzyme-linked immunosorbent assay) is a plate-based assay technique designed for detecting and quantifying soluble substances such as peptides, proteins, antibodies, and hormones.
CHAPTER 1 SEMESTER V - ROLE OF PEADIATRIC NURSE.pdfSachin Sharma
Pediatric nurses play a vital role in the health and well-being of children. Their responsibilities are wide-ranging, and their objectives can be categorized into several key areas:
1. Direct Patient Care:
Objective: Provide comprehensive and compassionate care to infants, children, and adolescents in various healthcare settings (hospitals, clinics, etc.).
This includes tasks like:
Monitoring vital signs and physical condition.
Administering medications and treatments.
Performing procedures as directed by doctors.
Assisting with daily living activities (bathing, feeding).
Providing emotional support and pain management.
2. Health Promotion and Education:
Objective: Promote healthy behaviors and educate children, families, and communities about preventive healthcare.
This includes tasks like:
Administering vaccinations.
Providing education on nutrition, hygiene, and development.
Offering breastfeeding and childbirth support.
Counseling families on safety and injury prevention.
3. Collaboration and Advocacy:
Objective: Collaborate effectively with doctors, social workers, therapists, and other healthcare professionals to ensure coordinated care for children.
Objective: Advocate for the rights and best interests of their patients, especially when children cannot speak for themselves.
This includes tasks like:
Communicating effectively with healthcare teams.
Identifying and addressing potential risks to child welfare.
Educating families about their child's condition and treatment options.
4. Professional Development and Research:
Objective: Stay up-to-date on the latest advancements in pediatric healthcare through continuing education and research.
Objective: Contribute to improving the quality of care for children by participating in research initiatives.
This includes tasks like:
Attending workshops and conferences on pediatric nursing.
Participating in clinical trials related to child health.
Implementing evidence-based practices into their daily routines.
By fulfilling these objectives, pediatric nurses play a crucial role in ensuring the optimal health and well-being of children throughout all stages of their development.
CHAPTER 1 SEMESTER V PREVENTIVE-PEDIATRICS.pdfSachin Sharma
This content provides an overview of preventive pediatrics. It defines preventive pediatrics as preventing disease and promoting children's physical, mental, and social well-being to achieve positive health. It discusses antenatal, postnatal, and social preventive pediatrics. It also covers various child health programs like immunization, breastfeeding, ICDS, and the roles of organizations like WHO, UNICEF, and nurses in preventive pediatrics.
Navigating the Health Insurance Market_ Understanding Trends and Options.pdfEnterprise Wired
From navigating policy options to staying informed about industry trends, this comprehensive guide explores everything you need to know about the health insurance market.
CRISPR-Cas9, a revolutionary gene-editing tool, holds immense potential to reshape medicine, agriculture, and our understanding of life. But like any powerful tool, it comes with ethical considerations.
Unveiling CRISPR: This naturally occurring bacterial defense system (crRNA & Cas9 protein) fights viruses. Scientists repurposed it for precise gene editing (correction, deletion, insertion) by targeting specific DNA sequences.
The Promise: CRISPR offers exciting possibilities:
Gene Therapy: Correcting genetic diseases like cystic fibrosis.
Agriculture: Engineering crops resistant to pests and harsh environments.
Research: Studying gene function to unlock new knowledge.
The Peril: Ethical concerns demand attention:
Off-target Effects: Unintended DNA edits can have unforeseen consequences.
Eugenics: Misusing CRISPR for designer babies raises social and ethical questions.
Equity: High costs could limit access to this potentially life-saving technology.
The Path Forward: Responsible development is crucial:
International Collaboration: Clear guidelines are needed for research and human trials.
Public Education: Open discussions ensure informed decisions about CRISPR.
Prioritize Safety and Ethics: Safety and ethical principles must be paramount.
CRISPR offers a powerful tool for a better future, but responsible development and addressing ethical concerns are essential. By prioritizing safety, fostering open dialogue, and ensuring equitable access, we can harness CRISPR's power for the benefit of all. (2998 characters)
We understand the unique challenges pickleball players face and are committed to helping you stay healthy and active. In this presentation, we’ll explore the three most common pickleball injuries and provide strategies for prevention and treatment.
Struggling with intense fears that disrupt your life? At Renew Life Hypnosis, we offer specialized hypnosis to overcome fear. Phobias are exaggerated fears, often stemming from past traumas or learned behaviors. Hypnotherapy addresses these deep-seated fears by accessing the subconscious mind, helping you change your reactions to phobic triggers. Our expert therapists guide you into a state of deep relaxation, allowing you to transform your responses and reduce anxiety. Experience increased confidence and freedom from phobias with our personalized approach. Ready to live a fear-free life? Visit us at Renew Life Hypnosis..
Navigating Challenges: Mental Health, Legislation, and the Prison System in B...Guillermo Rivera
This conference will delve into the intricate intersections between mental health, legal frameworks, and the prison system in Bolivia. It aims to provide a comprehensive overview of the current challenges faced by mental health professionals working within the legislative and correctional landscapes. Topics of discussion will include the prevalence and impact of mental health issues among the incarcerated population, the effectiveness of existing mental health policies and legislation, and potential reforms to enhance the mental health support system within prisons.
QA Paediatric dentistry department, Hospital Melaka 2020Azreen Aj
QA study - To improve the 6th monthly recall rate post-comprehensive dental treatment under general anaesthesia in paediatric dentistry department, Hospital Melaka
Immunity to Veterinary parasitic infections power point presentation
Aishwary presentation 1
1. Dr.APJ ABDUL KALAM TECHNICAL UNIVERSITY
(Formerly Uttar Pradesh Technical University ) LUCKNOW
SUBMITTED BY :-
AISHWARAY MUDGAL
M.PHARM Ist SEM
0211COL016
TOPIC :- ENZYME LINKED IMMUNOSORBENT ASSAY
Noida Institute of Engineering & Technology
( Pharmacy Institute )
SUBMITTED TO :-
Dr. Saumya Das
Associate Professor
Noida Institute of Engineering and
Technology (Pharmacy Institute),
Greater Noida.
2. ELISA INTRODUCTION :-
It is an important immunological method for detecting and measuring antigens and
antibodies. It is a plate based assay technique designed for detecting and quantifying
substance such as peptides, proteins, antibodies and hormone.
It is based on the same principle as that of radioimmunoassay.
The main difference is that of enzyme immunoassay the antigen of antibody is conjugated
to an enzyme rather than a radioactive isotope.The enzyme is then detected by its ability
to convert a colourless substance into coloured one.
Enzyme immunoassay have become very popular in view of their high sensitivity, safety,
economy and the simple instrument requirement.
3. SOME BASIC TERMS
SOLID PHASE :- Usually microtiter plate well having 96 wells.
ADSORPTION :- Process of having antigen or antibody ,diluted in buffer,so it attached to
solid phase on incubation.
WASHING :- Simple flooding and emptying of wells with buffer solution to separate
bound reagent from unbounded reagent in ELISA.
ANTIGEN :- Molecule entity responsible to produce antibody when introduced inside
body.
ANTIBODY :- Protein produced in response to antigen.
ENZYME CONJUGATE :- Enzyme irreversible attach to antibody (horse reddish peroxide
,alkaline phosphatase).
CHROMOGEN :- Chemical alters colour as a result of enzyme interaction with substrate
4. PRINCIPLE OF ELISA
ELISA is based on following observations:
1. Antigens and antibodies can attach to polystyrene plastic plates or other
solid phase support and maintain immunological capabilities.
2. Antigens and antibodies can be bonded to exam and the resulting complexes
are fully functional both immunologically as well as enzymatically
3. Specity of antigen antibody reaction.
5. TYPES OF ELISA
1. Sandwich ELISA
2. Indirect ELISA
3. Direct ELISA
4. Competitive ELISA
Double-Antibody-Sandwich method
In this technique the wells and depression in a polystyrene plate are first coated with an antibody and the
sample containing antigen is then added and allow to react with the bound antibody. The wells is washed and
the second enzyme linked specific antibody is added and allowed to react. This result is an antibody(with
enzyme)-Antigen antibody sandwich. Finally the enzyme substrate is added for reaction with the enzyme. The
rate of enzyme action is directly proportional to the Amount of test antigen.
6. Enzyme activity may be followed by a colour change which can
which can be inspected visually or measured by colourimeter
this method has been used to assay of Hepatitis B-antigen.
7. Indirect Elisa Method
The principle of this test can be illustrated by outlining its application for detection of anti-HIV-1 and
anti-HIV-2 antibodies in the patient serum the wells of the polystyrene microtitre plate are coated
with antigen. Test anti serum is added and allow to incubate. If the antibodies in the antiserum are
homologous they will bind to immobilized antigen. The well is then washed and enzyme –labelled
antihuman antibodies are added to the system which link with the antibody –antigen complexes.
ELISA is a simple and versatile technique. ELISA kits are commercially available for the detection of
anti HIV, Hepatitis B surface antigen and rotavirus. ELISA kits have also been develop for detecting
Hepatitis-A-virus (from stool), Haemophilus influenza antigens (from spinal fluid), Toxoplasma
antigens(from serum),Entamoeba histolytica antigen(from faeces), Escherichia coli enterotoxin(from
stools)etc.
8. The enzyme substrate is added the rate of its degradation is
associated with colour change proportional to the
concentration of antibody present in the test sample the colour
change can be measured visually and by colourimeter.
9. DIRECT ELISA :-
• It uses a primary labeled anti-body that react directly with
antigen .
• It can be performed with the antigen that is directly
immobilized on assay plate.
• Not widely used but common for immuno-histochemical
Staining of cells & tissues.
COMPETITIVE :-
• Antibody coated microwell.
• Serum antigen & labeled antigen added together ….. Competition.
• Ab-Ag enzyme complex bound is inversely related to the concentration of antigen present in sample.
• Increased serum antigen results in reduced binding of Ag-enzyme conjugated with the antibody
producing less enzyme activity & (yellow) color formation.
• Used to determine small molecules like T3,T4 & Progesterone.
10. APPICATION OF ELISA:-
Measuring hormone level in blood.
- HCG (pregnancy)
- LH (ovulation)
- TSH,T3,T4 (thyroid).
Detection of infection.
- Sexually transmitted agents (HIV)
- Hepatitis B & C.
Detection allergen in food.
Used for rapid testing.
Used in vaccine QC.
For detecting genetically modified organism.
11. REFRENCES :-
o https://www.berthold.com/microplate/washers
o https://pubmed.ncbi.nlm.nih.gov
o https://www.slideshare.net/sabaahmed56/elisa-ppt-53678570
o https://medlineplus.gov/ency/article/003332.htm
o S.R. Sankar, Text book of pharmaceutical analysis, Third edition, Page
no. 28.1-28.8.
