Strategies for analysing and improving the expression and quality of recombinant proteins made in mammalian cells

Abstract

The production of monoclonal antibodies and other recombinant proteins is one of the highest growth areas in the pharmaceutical industry. Mammalian cells are used to manufacture the majority of biotherapeutics, largely due to their ability to perform complex post-translational modifications. Although significant progress has been made recently in improving product yields and protein quality, many challenges still lie ahead to achieve consistently high yields while avoiding potentially damaging protein modifications. The present review first considers the strategies used to analyse and improve recombinant protein expression of industrial cell lines, with an emphasis on proteomic technologies. Next, cellular and environmental influences on protein production and quality are examined, and strategies for improvements in product yield and quality are reviewed. The analytical techniques required to detect these protein changes are also described, together with prospects for assay improvements.

Utilizing combinatorial engineering to develop Tie2 targeting antagonistic angiopoetin-2 ligands as candidates for anti-angiogenesis therapy

“…Similarly, a number of studies have shown significantly decreased stability of single domain ligands engineered in isolation from their stabilizing domains [50, 51]. Finally, low levels of recombinant expression have been reported for multidomain mammalian proteins, such as growth factor ligands and RTKs with clinical potential [52, 53]. …”

Tailoring recombinant protein quality by rational media design

“…The efficacy, potency and safety depend on the structural characteristics of the protein entity 11 , which explains the particular attention paid in biotherapeutic development to post-translational modifications that the expressed protein may undergo 1,12,14,[39][40][41] . In the particular case of monoclonal antibodies (mAbs), increased antibody-dependent cell-mediated cytotoxicity (ADCC) is one of the major objectives to increase the immune response of the human body and thus clinical efficacy 42 .…”

“…Most Cys are involved in disulfide bridges. Met oxidation has been reported to adversely affect both mAb structure and stability 39 . Various impacts of oxidation on biological activity, stability, and half-life of the therapeutic protein have been identified, and oxidationinduced aggregation may provoke immunogenicity reactions 68 .…”

Production of Recombinant Human Granulocyte Macrophage Colony- Stimulating Factor from Silkworm Bombyx mori Bm5 Cells

“…Thus, they are routinely used for the production of commercially valuable recombinant protein materials. However, the cost of human hormones used in the production of recombinant proteins using mammalian cell lines is a major problem (Jenkins et al, 2009).…”