Open AccessOptimization of BY ‐2 cell suspension culture medium for the production of a human antibody using a combination of fractional factorial designs and the response surface method
Author(s) -
Vasilev Nikolay,
Grömping Ulrike,
Lipperts Anja,
Raven Nicole,
Fischer Rainer,
Schillberg Stefan
Publication year - 2013
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12079
Subject(s) - fractional factorial design , factorial experiment , response surface methodology , suspension (topology) , nutrient , yield (engineering) , biology , antibody , suspension culture , food science , chromatography , plackett–burman design , central composite design , cell culture , chemistry , materials science , immunology , mathematics , composite material , ecology , statistics , genetics , homotopy , pure mathematics
Summary We have developed a strategy for the optimization of plant cell suspension culture media using a combination of fractional factorial designs ( FFD s) and response surface methodology ( RSM ). This sequential approach was applied to transformed tobacco BY ‐2 cells secreting a human antibody ( M 12) into the culture medium, in an effort to maximize yields. We found that the nutrients KNO 3 , NH 4 NO 3 and C aCl 2 and the hormones 2,4‐dichlorophenoxyacetic acid (2,4–D) and 6‐benzylaminopurine ( BAP ) had the most significant impact on antibody accumulation. The factorial screening revealed strong interactions within the nutrients group ( KNO 3 , NH 4 NO 3 and C aCl 2 ) and also individually between 2,4‐D and three other components ( KNO 3 , NH 4 NO 3 and BAP ). The RSM design resulted in a fivefold increase in the antibody concentration after 5 days and a twofold reduction in the packed cell volume ( PCV ). Longer cultivation in the optimized medium led to the further accumulation of antibody M12 in the culture medium (up to 107 μg/mL, day 10). Because the packed cell volume was reduced in the optimized medium, this enhanced the overall yield by 20‐fold (day 7) and 31‐fold (day 10) compared to the conventional MS medium.