earticle

영어

The kinetic properties required for sensitive and rapid sandwich ELISAs were determined rationally based on the kinetic model. And the screening conditions for isolation of target antibodies from a phage display library was designed based on simulation. The solid phase antibody are required to possess a sufficiently low koff value to minimize dissociation of the triple complex of the solid phase antibody, the antigen and the enzyme-conjugated antibody during the reaction in the solution of the conjugated antibody. Both the solid phase and the conjugated antibodies should possess high kon values to ensure a rapid reaction during a limited reaction time. However, with a decrease in koff value and with an increase in kon value, the appearance frequency in a repertory of antibody decreases1). Thus, the screening conditions were designed so that the most desirable antibodies would be isolated from a given diversity. Under the
designed conditions, antibodies with the required kinetic properties were isolated successfully. Furthermore, we showed that the best combination of both antibodies for sandwich ELISA can be determined based on the kon and koff values of the candidate antibodies. Our rational strategy would contribute to rapid development of sensitive ELISAs.