Cell Culture Processes in Monoclonal Antibody Production

Abstract

This chapter outlines cell culture processes in monoclonal antibody production. Cell culture process development starts with cell line generation and selection, followed by media and culture condition optimization in small‐scale systems, including 96‐well plate, shaker flasks, and bench‐scale bioreactors, for high throughput screening purposes. Once the process conditions are defined, the process is often transferred to the pilot scale to get scale‐up information and to produce toxicology materials and later to cGMP manufacturing for production of clinical material. As development of a commercial cell culture process for production of a biological product is completed at the laboratory and pilot scales, the commercialization process begins with process characterization, scale‐up, technology transfer, and validation for the manufacturing process. The strong demand of therapeutic antibody production, relatively modest titers, and reduction of cost of goods has resulted in a trend toward large‐scale manufacturing. Today, the largest biotech companies are using several hundred to 2,000 L scale bioreactors for early clinical production and 10,000‐25,000 L stirred tank bioreactors to produce commercial products.

Effects of passage number on growth and productivity of hybridoma secreting MRSA anti-PBP2a monoclonal antibodies

“…Such loss of production between initial and end-of-production cells might compromise regulatory approval and, in the worst-case scenario, could result in rejection of a particular cell line after months of wasted development effort (Barnes et al 2003). In addition to cell line stability, growth and metabolite characteristics affecting process robustness and scalability also need to be assessed (Li et al 2010). Despite the mAb tendency to lose productivity over a time, the limit in the number of passages that a given cell can be subjected to without impinging on its stability is singular to each cell line (Lee et al 1991).…”

“…Considering cost-effectiveness, it is important to have a cell line secreting proteins or antibodies of interest for development of diagnostics kits (Barnes et al 2003;Schmid et al 1990;Xin and Cutler 2006;Beckmann et al 2012;Kim et al 2011;Li et al 2010;Lee et al 1991). …”

Tailoring recombinant protein quality by rational media design

“…When antibodies began to enter the development pipelines, the biotech manufacturing changed over to fed-batch mainly due to productivity reasons. Antibody therapies may require large doses over a long period of time 33 . In that context, many companies have built large-scale facilities of working volumes ≥ 10,000 L. In the mid-2000, product titers of 5 g/L at the end of the production have become the industry standard 8 .…”

“…In that context, many companies have built large-scale facilities of working volumes ≥ 10,000 L. In the mid-2000, product titers of 5 g/L at the end of the production have become the industry standard 8 . Nowadays, biotechnology companies are reporting productivities as high as 10-13 g/L in a fed-batch culture of 2-3 weeks 33,34 .…”

“…These also called high-molecular-weight species (HMW) have to be limited to avoid immunogenic reactions in the injected patients. It is also important to keep in mind that the productivity of recombinant cell culture processes has substantially increased from 50 mg/L in batch to titers of 10 to 13 g/L nowadays in fed-batch culture of 2 to 3 weeks 33,34 . Moreover, the literature describes how cell culture conditions can be identified to control protein aggregation in CHO cell cultures 54 .…”