Syed Hussain Mir
University of Freiburg, Germany
Title: Generation of avian monoclonal antibody fragments for membrane protein crystallization.
Biography
Biography: Syed Hussain Mir
Abstract
Membrane proteins are challenging targets for crystallization and structure determination by X-ray crystallography. Antibody mediated crystallization has a major impact on the advancing structural and functional characterization of difficult membrane proteins1,2,3. More than 26 unique structures of membrane protein - antibody complexes have already been determined. An update of methods for generation of recombinant antibodies from hybridomas and their production in E. coli was recently published (Mir et al. 2015)3. The limited availability of suitable hybridoma cell lines due to low immunogenicity of therapeutically important human membrane proteins in mice has impeded the high-throughput application of this approach. Here we show an efficient method to obtain high affinity binders against difficult targets by phage display exploiting the avian immune system. The recombinant chicken antibodies were generated against Na+/H+ transporter (STNhaA) and used for its structural characterization. The strategy of avian immune phage display libraries provides fast access to versatile tools for structural and functional studies and in general paves the way to generate versatile tools for research, diagnostics and therapeutics targeting membrane proteins and is of special interest for antigens highly conserved in mammals.