Functional characterization of two anti‐estradiol antibodies as deduced from modelling and site‐directed mutagenesis experiments

• DOI: 10.1002/jmr.525
• Publication Date: 2002-08-25T20:23:10Z
• Authors: Florence Bettsworth; Céline Monnet; Bénédicte Watelet; Nicole Battail‐Po...; Bernard Gilquin; Michel Jolivet; André Menez; Michel Arnaud; Frédéric Ducancel
• Keywords: Immunogen; Bovine serum albumin; Site-directed mutagenesis

Monoclonal antibodies are now widely used to measure the concentration of steroid hormones in human serum samples. The great development molecular engineering techniques over past 10 years has made possible improvement specificity and/or sensitivity selected antibodies. We have obtained two monoclonal antibodies, 17E12E5 and 10G6D6, using estradiol-6-ethyl methoxy carbonyl (EMC)-bovine albumin (BSA) as immunogen. To tentatively improve their affinities for natural estradiol, we initiated structural functional studies. For this purpose, cloned sequenced genes encoding variable fragments each antibody. Single chain (scFv) were produced into periplasmic space E. coli pLIP6 expression vector. Mapping structures both was by combination modelling mutational analyses together with cross-reaction binding pockets described models estradiol complexed 10G6D6 proposed.