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Tailoring translational strength using Kozak sequence variants improves bispecific antibody assembly and reduces product‐related impurities in CHO cells
Author(s) -
Blanco Noelia,
Williams Ambrose J.,
Tang Danming,
Zhan Dejin,
Misaghi Shahram,
Kelley Robert F.,
Simmons Laura C.
Publication year - 2020
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27347
Subject(s) - chinese hamster ovary cell , sequence (biology) , heterologous , recombinant dna , antibody , protein engineering , transfection , computational biology , cell culture , chemistry , microbiology and biotechnology , biology , enzyme , biochemistry , genetics , gene
Optimal production of bispecific antibodies (bsAb) requires efficient and tailored co‐expression and assembly of two distinct heavy and two distinct light chains. Here, we describe a novel technology to modulate the translational strength of antibody chains via Kozak sequence variants to produce bsAb in a single cell line. In this study, we designed and screened a large Kozak sequence library to identify 10 independent variants that can modulate protein expression levels from approximately 0.2 to 1.3‐fold compared with the wild‐type sequence in transient transfection. We used a combination of several of these variants, covering a wide range of translational strength, to develop stable single cell Chinese hamster ovary bispecific cell lines and compared the results with those obtained from the wild‐type sequence. A significant increase in bispecific antibody assembly with a concomitant reduction in the level of product‐related impurities was observed. Our findings suggest that for production of bsAb it can be advantageous to modify translational strength for selected protein chains to improve overall yield and product quality. By extension, tuning of translational strength can also be applied to improving the production of a wide variety of heterologous proteins.