Monoclonal Antibody Process Development Using Medium Concentrates

Abstract

A fed-batch process using concentrated medium was evaluated for its ability to improve cell culture longevity and final monoclonal antibody (MAb) titers for two monoclonal antibody producing cell lines. It was found to result in up to 7-fold increases in final antibody titers compared to batch culture controls. Although the development cell line specific fed-batch protocols is critical to the development of cost-efficient large-scale production processes, the use of complete medium concentrates provided us with a quick and simple method for producing large quantities of antibodies in the early stages of process development, thus accelerating early work on purification process development, analytical development, biochemical characterization, and safety studies. Insights gained from the concentrated medium fed-batch approach were valuable for the development of refined, cell line specific feeding strategies yielding final MAb titers on the order of 1-2 g/L. Process development data on the effects of inhibitory growth byproducts, medium osmolarity, and the mode of nutrient feed addition on culture longevity and MAb production and information on culture metabolic behavior were successfully incorporated in the development of the optimized fed-batch protocols.

Introduction to animal cell culture technology—past, present and future

“…Then the early nineties were marked by the development of recombinant GS-NS0 technology (by Celltech) leading a few years later to the possibility of producing 1-2 g/l of monoclonal antibodies in a fed batch process (Bibila et al 1994). Today we see antibody titers at an industrial scale of 5 g/l and more (Birch 2005).…”

Study on Interactions between Fed-Batch and Batch Operating Parameters for the Development of Monoclonal Antibody Fed-Batch using Design of Experiments

“…Feeding medium concentrates has been reported to dramatically increase cell densities and final product titers in recombinant CHO [17] and NS0 cell cultures [3]. Figure 3 shows the consumption and production profiles for the most important biochemical components in the growth medium namely Glucose, Lactate, Glutamine and Ammonia.…”

“…However, for the choice of the appropriate cell line, certain key criteria have to be considered which include: the ability to provide a close to human glycosylation pattern, the capability to produce high mAb concentrations in the chosen production system, the ability to consistently produce a product of uniform characteristics (stability), and the speed with which a high yielding cell line can be obtained [1,2]. NS0, a murine myeloma cell line, has been adopted by a number of biotechnology companies for the expression of therapeutic antibodies [3][4][5][6][7]. The successful use of NS0 cells in several fusion, transfection, selection and production approaches are among the few properties of this cell line that make it a perfect candidate for the expression of the desired product [8].…”

“…Fed batch cultures have been developed with the objective of achieving maximal increase in the culture viable cell concentrations and prolonging the culture lifetime for increased product concentrations [3,5,[11][12][13][14][15] Strategies such as development of the feeding solutions, feeding rate control, maintaining low residual glucose and/or glutamine concentrations, employing dialysis membranes to remove low molecular weight components from the culture, feeding based on Oxygen UptakeHence, based on the hypothesis that none of the factors affecting the production process can really be independent, fractional factorial design using design expert software was carried out to find the interaction between the batch (Dissolved Oxygen, Temperature and Seeding Density) and fed-batch (post-feed Glucose concentration) process parameters. The usage of design of experiments also facilitates shorter development time, hence more cost effective.…”

Discrimination among IgG1-κ monoclonal antibodies produced by two cell lines using charge state distributions in nanoESI-TOF mass spectra

“…Humanized IgG1-monoclonal antibodies (mAbs) were produced in two cultivation systems, CHO [16] and GS-NS0 [17] cells. A four-step purification platform involving solubility, size separation, charge separation, and filtering were used.…”