Biochemistry & Analytical Biochemistry

Generation of avian monoclonal antibody fragments for membrane protein crystallization

International Conference on Biochemistry

October 10-12, 2016 Kuala Lumpur, Malaysia

Syed Hussain Mir, Christopher Lentes, Christophe Wirth, Anika Fippel and Carola Hunte

University of Freiburg, Germany
University of Kashmir, India

Posters & Accepted Abstracts: Biochem Anal Biochem

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 proteins 1, 2 and 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. 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 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.

Biography :

Email: syed.mir@biochemie.uni-freiburg.de