Radioimmunoassay and ELISA

1. RADIOIMMUNOASSAY(RIA)
Rounak Bhattacharya
M-Pharm(Pharmaceutics), 1st year(1st Sem)
University Roll no. 18920321007
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Content:-
Introduction
Definition
Principle
Merits and Demerits of RIA
Detailed procedure in RIA
ELISA
Instrumentation
Applications
Conclusion
Reference

3. Introduction:-
 Immunoassays are bioanalytical methods that use the specificity of an
antigen-antibody reaction to detect and quantify target molecules in
biological samples. These methods are frequently used in clinical
diagnostics, drug discovery, drug monitoring, and food testing.
 Radioimmunoassay(RIA) was discovered by Berson and Yellow
in 1959 for the quantitative measurement of insulin in human
plasma.
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Definition:-

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Principle:-

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Merits and Demerits
Merits:-
 Useful for measuring trace amounts of antibodies, hormones, proteins, vitamin etc.
 Required very small amount of sample(5-50) picomoles/ml
 Simple, rapid, accurate highly specific and highly sensitive method.
Demerits:-
 Required special counting equipment.
 Required skilled personnel and is an expensive process.
 Laboratories required special license to handle radioactive materials.
 Required special arrangements for storage of radioactive material and radioactive waste
disposal.

7. Procedure:-
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Radio
activity
Conc. Of cold antigen

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ELISA(ENZYME LINKED
SORBENT ASSAY)
Antigen
coated wall
Specific
antibody
binds to
antigen
Enzyme linked
antibody binds to
specific antibody
Substrate is added and
converted by enzyme into
coloured product, the rate
of colour formation is
proportional to the amount
of specific antibody
Wash Wash Wash

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Procedure:-

10. Instrumentation:-
 Centrifuge:-
Swing bucket rotor:- 1200-2500 rpm
Fixed angle head rotor:- 3500-4000 rpm
 Radio active counter:-
Gama counter :- Which is used for gamma energy emitting isotope
Scintillation counter:- It is used for beta emitting isotope.
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 Analysis of hormone, vitamin:- ACTH, FSH,Insuline, Vitamin B12
 Therapeutic drug monitoring:- Morphine, Digoxin
 Diagnostic procedure for detecting infection:- HIV, Hepatitis A
 Non clinical application:- Food processing industry
Application:-

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Conclusion:-
Thus , it can be concluded that at present, Radioimmunoassay(RIA) is the most widely used highly sensitive
analytical technique available for measuring small concentrations of biological and pharmacological
substances in different samples. The most important drawback was the health threat due to the application of
radioactive elements in this technique. Subsequently, new alternatives were developed based on using
nonradioactive substances as labels. Although RIA is somewhat an outdated technique, its principles are
known to be still applied to other systems that operate based on specific binding reagents.

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Reference:-
 Van Vunakis, H., 1980. [10] Radioimmunoassay: An overview. Methods in
enzymology, 70, pp.201-209.
 Block, E., 1978. An overview of radioimmunoassay testing and a look at the
future.
 Too, H.P. and Maggio, J.E., 1991. Radioimmunoassay of tachykinins.
In Methods in neurosciences (Vol. 6, pp. 232-247). Academic Press.
 Dr.S. Ravi Sankar, "Pharmaceutical analysis” 267 A/3, West Car Street,
Tirunelveli - 627 006, India, pp.33.1-33.07,2010
 http://www.authorstream.com/Presentation/dodda_sri-1396092-ria-ppt
(Accessed on 02.03.22)
 https://www.slideshare.net/JSwaminathan93455166/radio-immuno-assay-
76545400 (Accessed on 02.03.22)

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