Cell culture compositions and methods for polypeptide production
A composition, cell technology, applied in biochemical equipment and methods, cell culture media, cell culture active agents, etc.
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Embodiment 1
[0384] Example 1: Color intensity exhibited in formulations containing antibodies isolated from antibody-producing cells
[0385] A CHO cell line capable of producing an IgG1 monoclonal antibody (anti-β7) was grown in chemically undefined medium containing peptone. The isolated antibody was purified and color was evaluated using the standard "Clarity, Opalescence and Coloration" (COC) test (Council of Europe. European Pharmacopoeia., 2008, 7th edition, p. 22). Briefly, the COC test was carried out using the same flat-bottomed colorless transparent neutral glass tube with an inner diameter of 15 mm to 25 mm. A 150 g / L protein solution prepared from purified concentrated cell culture fluid containing secreted IgGl monoclonal antibodies was added to the tube to a depth of 40 mm. The tube containing the antibody solution was compared to 9 reference tubes each containing a reference solution from B1 (darkest) to B9 (lightest) by viewing perpendicular to a white background in diffu...
Embodiment 2
[0389] Example 2: Reducing the Color Intensity of Antibodies Isolated from Antibody-Producing Cell Lines by Altering Specific Components in the Cell Culture Medium
[0390] Basal Medium 1 and Feed Medium 2 were reformulated to contain reduced concentrations of several nutrients and used in cell culture experiments to determine if this reformulated medium could reduce the production of isolated monoclonal IgG1 antibodies from CHO cell lines (anti-β7) color intensity. Briefly, basal CDM (medium 3) and fed CDM (medium 4) solutions were prepared by combining the ingredients into a custom blended powder, dissolving the powder in water, and adjusting to ensure optimal cell growth. Final pH and osmolarity. Basal Medium 3 and Feed Medium 4 each had more than 20 components, of which the components of interest are listed in Table B. Some media components such as glucose are not combined in the mixed powder, but added separately during the media preparation process. Similarly, media c...
Embodiment 3
[0397] Example 3: Reduction of Color Intensity of Antibodies Isolated from Antibody-Producing Cell Lines by Altering Vitamin B Levels in Cell Culture Media
[0398] To determine the effect of changing components in basal medium 3 and feed medium 4 on the color intensity of isolated antibodies, the levels of vitamins B2, B6, B9 and B12 were varied while the levels of other medium components were kept constant. The medium was prepared as described in Example 2. Media components altered for this study were not included in the mixed powders but were added separately at appropriate levels during media formulation. Basal Medium 5 was formulated to have similar reduced vitamin levels as Basal Medium 3 and all other ingredients similar to Basal Medium 1 (Table C). For the production of monoclonal IgG1 antibodies (anti-β7) from CHO cells, cell cultures were fed basal medium 5 for the initial growth phase and fed with either feed medium 2 or feed medium 4 on days 3, 6, and 9. To culti...
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