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Radioimmunoassay by Group 7

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27 views2 pages

Radioimmunoassay by Group 7

Uploaded by

Ngetwa Wiryboy st
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Radio Immuno Assay (RIA)

Radio immuno assay can be used to determine the concentration of


antigens like harmones, serum protein, drug and vitamins in
biological fluids based on antigen antibody reaction and measuring
the radioactivity of resultant labelled antigen antibody complex.
This technique was developed by berson and yalow in 1960 they got
nobel prize in 1977
This technique is very sensitive that it can detect 0.001ug/ml
concentration of antigens.
Principle:
The principle of RIA is based on antigen antibody reaction in which
the radio labelled antigen competes with the endogenous antigen for
the limited sites of the specific antibodies against the same antigen
the radio labelled antigen should have similar biological activity and
immunogenicity like that native antigen.
Antibody+ Antigen+ labelled antigen: Antigenfi Antibody + labelled
antigen - Antibody + Antigen+ labelled antigen.
Procedure:
1. To carry out RIA procedure one require
Anti A Antibodies: which bind to antigens in the sample which
are prepared by hydrobloma technique
• Radiolabelled A Antigen: which some part is made up of Radio
isotopes so that they can emit raditation like alpha or beta,
heavy isotopes for H2
• Unlabelled antigens
2. Take microtile plate and coat with the antibody
3. Then add Radio labelled antigens in excess amounts so that no
antigen bounding sites left free in which some Radio labelled
antigens are not bounded to the antibodies are removed by washing.
4. Hence the radioactivity of antigens -antibody is 100%
ft. Then add known quantity of unlabelled antigen to the well which
creates the competition between labelled and unlabelled antigens
for binding with the antibody. This competition is proportional to the
concentration of unlabelled antigens added to the well.
6. Some labelled antigens will be removed from the antibody which
are replaced by unlabelled antigen and labelled and the radioactivity
of antibodies is not 100%
7. Repeat the procedure by increasing the concentration of
unlabelled antigens results in lesser radioactivity.
8. Plot a graph between concentration of unlabelled antigen and
Radioactivity of labelled antigen -antibody complex which gives a
linear graph.
9. Repeat the procedure by taking sample in place of unlabelled
antigen.
10. The antigens which are present in the sample will be compete
with the labelled antigen and forms a complex.
11. The value of Radioactivity will give the concentration of antigens
present in the sample from the linear graph.
Applications of RIA:
1. RIA has been used to assay plasma levels of most harmones,
Insulin in human plasma, betafi HCG in females, vasopressin
2. Digitoxin or digoxin in patients receiving these drugs

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