Protein-protein interactions are often modelled by antibody-antigen interactions and the understanding of molecular interactions underling antibody specificity and affinity are still to be resolved. \Ve have developed a mode! system of high affinity monoclonal antibodies specific for Hen Egg Lyso/yme (HEL). The structure of one of our antibodies HyHEL 10 complexed with H E I. has been defined by X-ray crystallography, and provide a definition of the st uctural epitope recognized by this antibody. The definition and the understanding of the interaction of this antibody with HEL require complementary sei of data, such as the definition of the residues contribul ing the energy of t ho interaction and defining the energetic epitope. Therefore the use of natural mutants or site directed mutants of both the antigen and the antibody is the too! of choice to answer these questions. HEL was actually used as prototypic antigen because of the numerous natural variants available, and we have designed and expressed around .'iO mutants of HEL in the methylatropic yeast pichia pastoris. 'I hese mutants obtained bv replacing residues near or at the interface of HyHEL 1ÜTIEI. complex with Ala, will be used to study the interaction with HyHKL 10 and also with two related antibodies of close specificity. HyHKL H and HyHEL 2fi. The sensitivity <>f these related antibodies to the mutants will also shed light on the contribution of antibody residues in the interaction, by correlating the sensitivities to the primary sequence of these diidtibodies.
|Publication status||Published - 1997|
ASJC Scopus subject areas
- Molecular Biology