Comparative Investigations of Codon Engineered Anti-EGFR Antibody to Enhance Protein Expression in Mammalian Host

Author(s): *Sharvari Hemendra Joshi, Archana Krishnan and Kiran Mangaonkar

Abstract: Monoclonal antibodies are extensively used in biomedical research, in diagnosis of diseases, and in the treatment of such diseases as infections and cancers. Molecular cloning and expression in mammalian host is the most efficient method for the expression of therapeutic monoclonal antibodies. Production of biotherapeutics like monoclonal antibodies in the mammalian host requires optimization of complex molecular parameters to accomplish desired expression such as codon optimization. Codon optimization is a competent gene engineering approach for effective protein expression. Gene designing enables exploitation of degeneracy of genetic code to boost target recombinant protein expression. The present work describes comparative analysis of codon optimization from two different sources. Trastuzumab genes were codon optimized by using GeneArt (Thermo fisher scientific) codon optimization services as well as in-house codon optimization algorithm. Trastuzumab genes were cloned and expressed in pBG-SVII(pgk) mammalian expression vector. In-house codon optimized Trastuzumab demonstrated proficient results in pBG-SVII(pgk) mammalian expression vector when compared to GeneArt codon optimization services (Thermo fisher scientific) codon optimization. Current study reveals that in-house codon optimization algorithm is advantageous to achieve higher level of monoclonal antibody expression in mammalian host.

PAGES: 787-795  |  31 VIEWS  48 DOWNLOADS