Determination of dissociation constants of Anti-ALP monoclonal antibodies by an ELIS A - based method

摘要

BACKGROUND: Dissociation constant (Kd) is of major significance in immunoassay. Since affinity may be influenced by the immunoassay methodology it is important to determine this parameter under the condition of the assay used.OBJECTIVE: To employ a rapid and simple ELIS A- based method for measuring dissociation constants of two Alkaline Phos-phatase (ALP) specific MAbs (A_1G_8F_7 and A_1G_9G_3) established in our laboratory.METHODS: A simple and rapid method on enzyme- linked immunosorbent assay (ELISA) was developed for measuring the dissociation constant of antibody - antigen reactions.In this method the ability of increasing amounts of antigen to bind to antibody measured in the fluid phase.