Efficient and effective supplement screening for the development of chemically defined media in cell culture

摘要

The present invention relates to methods of selecting and developing a chemically defined media (“CDM”) for use in the manufacture of biological products. In particular, the present invention is directed to screening methods to determine cell culture technique media supplement blends with enhanced performance characteristics. The present invention is also directed to identifying CDM supplement blends that demonstrate significant increases in harvest titer and/or viable cell density.

主权项

1. A high-throughput method of screening cell culture media supplements to determine a concentration of a supplement combination capable of inducing an increase in peak viable cell density in a cell culture, the method comprising:
a. supplementing a plurality of individual volumes of chemically defined media with a supplement combination comprising L-isoleucine (0.23-0.29 g/L); L-leucine (0.24-0.31 g/L); L-lysine (0.47-0.37 g/L); L-methionine (0.08-0.10 g/L); L-phenylalanine (0.15-0.19 g/L); L-threonine (0.21-0.26 g/L); L-tyrosine (0.15-0.19 g/L); L-valine (0.23-0.30 g/L); L-arginine (0.53-0.68 g/L); L-histidine*HCl*H2O (0.18-0.23 g/L); L-tryptophan (0.08-0.10 g/L); Calcium pantothenate (0.13-0.17 g/L); Cupric sulfate (0.02 g/L); a-cyclodextrin (0.14-0.18 g/L); P-cyclodextrin (0.14-0.18 g/L); Folate (0.05-0.06 g/L); Manganese chloride (5.91×10−5-7.59×10−5 g/L); Potassium phosphate (0.09-0.12 g/L); Thymidine (1.75×le 2.25×10−3 g/L); i-inositol (0.09-0.12 g/L); and Zinc chloride (1.82×10−3 2.33×10−3 g/L; b. inoculating the individual volumes with a cell line of interest; c. culturing the cell line of interest in the individual volumes; d. measuring peak viable cell density of the cultured cell line in the individual volumes; and e. performing Analysis of Variance statistical analysis of the measurements of peak viable cell density; wherein the results of the statistical analysis indicate the concentration of a supplement combination capable of inducing an increase in peak viable cell density in a cell culture.