Degradation of recombinant proteins by Chinese hamster ovary host cell proteases is prevented by matriptase-1 knockout

摘要: An increasing number of nonantibody format proteins are entering clinical development. However, one the major hurdles for production glycoproteins is host cell-related proteolytic degradation, which can drastically impact developability and timelines pipeline projects. Chinese hamster ovary (CHO) cells preferred recombinant therapeutic proteins. Using protease inhibitors, transcriptomics, genetic knockdowns, we have identified, out >700 known proteases in rodents, matriptase-1 as involved degradation expressed CHO-K1 cells. Subsequently, was deleted using "transcription activator-like effector nucleases" (TALENs) well zinc-finger nucleases (ZFNs). This resulted a superior matriptase (KO) cell line with strongly reduced or no activity toward panel recombinantly The KO evaluated spike-in experiments showed little incubated culture supernatant derived from effect confirmed when same were line. In summary, combination novel engineering tools, next-generation sequencing screening methods, recently published genome has enabled development this CHO capable improving expression yields intact