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Radioisotopes RIA and EL-27794
Radioisotopes RIA and EL-27794
Objectives
To explain the principle of radioactivity and its various types of
decay.
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http://images.tutorvista.com/cms/images/38/nuclear-decay-equations1.png
http://www.buzzle.com/images/diagrams/gamma-decay.jpg
Gamma ray has no mass and no charge, therefore penetration
power is maximum.
Hence Gamma rays are widely used for treatment of cancer cases
http://www.frankswebspace.org.uk/ScienceAndMaths/physics/physicsGCE/images/penetration.jp
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Applications of Radioactivity
Research
Diagnostic application
Therapeutic application
Applications of Radioactivity in
Research
Since isotopes of an element have identical chemical reactions, when a
radiolabeled compound is administered, these molecules are
metabolized by the body similar to the normal molecules. This is called
“Tracer technique”.
Almost all the biochemical pathways were studied by using tracers. Eg;
14 C – isotope labeled metabolites
PET scan with a series of images tells us about the functional status
of an organ. PET scan is widely used in oncology with 18 F as a tracer.
Radioimmunoassays (RIA)
Applications of Radioactivity in
Treatment
131I is used for the treatment of hyperthyroidism(Graves’
disease) and thyroid cancers.
Radioimmunoassay (RIA)
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Radioimmunoassay
The principle of RIA involves competitive binding of
radiolabeled antigen and unlabeled antigen to a high-
affinity antibody.
Step 1:
The labeled antigen is mixed with antibody at a concentration
that saturates the antigen-binding sites of the antibody.
Step 2:
Test samples of unlabeled antigen of unknown concentration
are added.
The antibody does not distinguish labeled from unlabeled
antigen, so the two kinds of antigen compete for available
binding sites on the antibody.
As the concentration of unlabeled antigen increases, more
labeled antigen will be displaced from the binding sites.
Step 3:
To determine the amount of labeled antigen bound, the Ag-Ab
complex is precipitated and the radioactivity in the precipitate is
measured.
Step 4:
A standard curve can be generated using unlabeled antigen
samples of known concentration (in place of the test sample),
and from this plot the amount of antigen in the test mixture may
be precisely determined.
Radioimmunoassay
These assays approach the sensitivity of RIAs and have the advantage
of being safer and less costly
Overview of steps in ELISA
Coating:
Polystyrene plate is treated with a solution of
either antigen or antibody
Blocking:
An unrelated protein based solution is used to
cover all unbound sites on the plate
Antibody-antigen interaction:
Enzyme conjugated antibody specifically binds
to target antigen or antibody
Detection:
Substrate specific for the enzyme is added and the
intensity of the color produced by the enzyme substrate reaction
is measured by spectrophotometry
Types of ELISA
ELISA can be used to detect qualitatively or quantitatively
the presence of antibody or antigen.
Types of ELISA:
Direct ELISA
Indirect ELISA
Sandwich ELISA
Direct ELISA
An antigen coated to a multiwell plate is detected
by an antibody that has been directly conjugated
to an enzyme.
Advantage:
It is performed faster since fewer steps are
Involved.
Disadvantage:
Less signal amplification and hence lower sensitivity
Indirect ELISA
Antigen coated to a polystyrene multiwell plate is
detected in two stages