Staphylococcal Surface Display in Combinatorial Protein Engineering and Epitope Mapping of Antibodies
Nina Kronqvist, Magdalena Malm, Johan Rockberg, Barbara Hjelm, Mathias Uhlen, Stefan Stahl and John Lofblom
Affiliation: School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, SE-106 91 Stockholm, Sweden.
Keywords: Affibody molecules, antibody engineering, bacterial display, cell surface display, combinatorial protein engineering, epitope mapping, staphylococcal surface display, yeast display, ENGINEERING, proteins, life science, Enzymatic, catalysis, amino acids, recombinant, polynucleotides, genotype, antibody clones, non-immunoglobulin, flow-cytometric analysis, optimization, staphylococcal, Saccharomyces cerevisiae, Gram-negative bacteria, E. coli, S. cerevisiae, fluorescein, –, biotin, hapten, screening, immune antibody, HIV antigen, IgG-binding, cell wall, propeptide, lactamase, metal-binding, cellulose-binding, domain, optimized display vector, SPR technology, multivalent targets, osmotic stabilizer, electroporation buffer, electroporation, frequency, growth-rate, monoclonal antibodies, rheumatoid arthri-tis
The field of combinatorial protein engineering for generation of new affinity proteins started in the mid 80s by the development of phage display. Although phage display is a prime example of a simple yet highly efficient method, manifested by still being the standard technique 25 years later, new alternative technologies are available today. One of the more successful new display technologies is cell display. Here we review the field of cell display for directed evolution purposes, with focus on a recently developed method employing Gram-positive staphylococci as display host. Patents on the most commonly used cell display systems and on different modifications as well as specific applications of these systems are also included. General strategies for selection of new affinity proteins from cell-displayed libraries are discussed, with detailed examples mainly from studies on the staphylococcal display system. In addition, strategies for characterization of recombinant proteins on the staphylococcal cell surface, with an emphasis on an approach for epitope mapping of antibodies, are included.
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