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For several decades, Escherichia coli has been considered as the most important host for recombinant protein production because of rapid growth, easy gene manipulation, easy scale-up etc. In addition, recent developments of various protein display system have allowed E. colI
to be an useful tool in protein engineering. Previously we have developed new protein display system, Anchored Periplasmic Expression (APEx) system, in which protein is anchored on the inner membrane of E. coli via lipoprotein fusion. Combination of flow cytometry (FC) and APEx system make it possible to do high throughput screening of protein variant with odified/improved properties such as different specificity, maturated affinity, enhanced stability etc.
Recently, based on APEx system, we have developed new 2-hybrid system (APEx 2-hybrid) in which two interacting proteins are coexpressed and is suitable for proteomic study as well as high throughput screening for protein engineering. Various applications of APEx and
APEx 2-hybrid systems toward antibody engineering (affinity maturation of antibody fragments and full-length IgG, development of intrabody etc.) will be presented.