Structural and Functional Characterization of a Codon Optimized Coagulation Factor VIII

Background. Production of recombinant factor VIII (FVIII) is challenging due to its low expression. Previously, it was shown that codon optimization of a B domain-deleted (BDD) FVIII resulted in its increased expression (Ward et al, Blood 2011; 117: 798-807). However, in some cases, synonymous mutations are known to affect the protein's post-translation modifications, conformation, fidelity of amino acid sequence, and functions. Thus, for each particular codon optimization of a given protein, confirmation of its biochemical characteristics is necessary. Recently, we established conditions for expression and purification of a codon optimized BDD-FVIII (CO), in parallel, testing the BDD-FVIII (WT) expressed from the wild-type cDNA sequence (Shestopal et al, ISTH-2015 meeting, Abstract PO196-WED). In present work, we verified if the characteristics of the CO remain unchanged upon modification of its coding sequence.