Kinetic Immunoassay*
The
typical antibody-antigen immunoassay technique uses
a labeled antibody (e.g. radioactive iodine, fluorescent
dye, etc.) to detect a specific antigen in an analyte
(the material to be analyzed). The antigen to be analyzed
is introduced into a solution of antibody specific
to antigen. While the antibody-antigen reaction is
very specific, reaction rates are typically low. Moreover,
tradition immunoassay techniques require that the
antibody-antigen reaction be allowed to proceed to
equilibrium. The result is that typical laboratory
immunoassay procedures are quite time consuming and
hence not convenient for field studies or routine
work.
Neogen
manufactures test kits for field use that are applicable
to the measurement of many chemicals; some of these
kits are based upon antibody-antigen reactions. It
is a priority in such immunoassays to shorten assay
time to increase the assay throughput. One way to
accomplish this objective is to measure data kinetically
over
a short incubation period (using a non-equilibrium
model) to speed the analysis. However, such methods
are complicated by the need to have detailed knowledge
of the antibody-antigen reaction kinetics. Thus, for
a given antibody-antigen pair, a kinetic model is
needed that accurately predicts the progress of the
reaction with time.
The project objective is to find a suitable model
for a given antibody-antigen pair.
Such
a model may be based upon biophysical parameters of
the antibody-antigen pair or they may be empirically
based. The former has the advantage of being able
to provide more detailed information about the antigen;
however, at the cost of greater analysis complexity.
On the other hand, an empirical model is simpler and
easier to implement but suffers from the disadvantage
of providing less information.
The project objective is to find a suitable kinetic
model and quantify its accuracy.
The
ideally completed project deliverable would be a suitable
kinetic model, either empirical or biophysical, and
a procedure for interpreting the reaction data to
find the amount of antigen in the analyte.
*This
summary was prepared by R.E. Svetic with the participation
of P. S. Satoh and J.M. Madden of Neogen Corporation.
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