Radio Immuno Assay

1. PRESENTED BY-
MR. DEBASHIS PUROHIT
M.PHARM,1st SEMESTER
(INDUSTRIALPHARMACY)
DATE-17/08/2017
SCHOOL OF PHARMACEUTICALSCIENCES,
SIKSHA ‘O’ ANUSANDHAN UNIVERSITY,BHUBANESWAR
TOPIC-THEORY AND APPLICATION OF
RADIO IMMUNO ASSAY(RIA)

2. Introduction[1] -
RIA(Radio Immuno Assay)?
 Immuno-It refers to an immune response which
causes the body to generate antibodies.
 Assay-It refers to a test.
 Immuno Assay-A test which uses immunocomplexing
when antigens and antibodies are brought together.

3. The technique was introduced in1960 by
Berson and Yalow as an assay for the
concentration of insulin in plasma.
It represented for first time in detection of
hormone levels in blood by invitro assay.

4. PRINCIPLE [1] -
Radioimmunoassay (RIA) involves the separation
of a protein (from a mixture) using the specificity
of antibody - antigen binding and quantitation using
radioactivity.
It uses Antigens and Antibodies for an immune
reaction.
This technique has revolutionary research and
clinical research in many areas like blood
banking,diagnosis of allergy etc.

5. THEORY[2] -
 Antigens-An antigen is a substance with the ability
to induce an immunological response. They
typically enter the body from an infection. They are
recognized at their antigenic determinant by B cells
or by the T cell receptor on T cells.Proteins or
glycoproteins make the best antigens because they
are the best at stimulating antigen recognition
molecules.

6. • Antibodies-Antibodies are produced by the B-
lymphocytes. They are glycoproteins
belonging to the "immunoglobulin supergene
family" that are produced in response to a
foreign substance in the body.

7. • Ag + Ag* + Ab -> AgAb + Ag*Ab + Ag + Ab*
• Unbound Ag* and Ag washed out Radioactivity
of bound residue measured Ligand conc. is
inversely related to radioactivity
• [Ag : ligand to be measured ; Ag* radiolabelled
ligand]

9. REQUIREMENTS OF RIA [2] -
Preparation and characterisation of the
Antigen(Ligand to be analyzed).
Radio labelling of the Antigen.
Preparation of the specific antibody.

10. ADVANTAGES[3] -
Highly specific
High sensitivity
It has become a major tool in clinical labrotory
where it is assayed.
Using Antigens of higher affinity, it is possible
to detect few picograms(10-12 ) of Antigen in
the tube.

11. DISADVANTAGES[3] -
The main drawbacks to radioimmunoassay are
the expense and hazards if preparing and
handling the radioactive antigen.
Labs requires special licence to handle radio
active materials.
Requires special arrangements for
requisition,storage of radio active material
and radio active waste disposal.

12. APPLICATIONS[3] -
Analysis of hormones,metabolites.Eg.
Testosterone,prostaglandin,insulin etc.
TDM i.e-Barbiturates,morphine etc.
Dignostic purpose for detecting infections.i.e. HIV.
Detection of Narcotic drugs.
Thyroid testing.
Diagnosis and treatment of Peptic ulcer.
Research with Neurotransmitters.

13. REFERENCE-
1-Kenneth A. Connors,A Textbook of Pharmaceutical Analysis,
Edition 3rd,pg no-556-562.
2-Edward j. Motica,L.C. Virchow,Europian Journal of
Pharmaceutical Science, A Historical Perspective on
Evidence-Based Immunology, 2016,pg no-353-364.
3-S.M. Glick, C.W. Rubenstein,Journal of Biomedical
Science, Nature of immunological responses and antigen-
antibody interaction,1967,pgno-844-856.