Two-dimensional electrophoresis as a tool for control of quality and consistency in production systems using animal cells. Two-dimensional electrophoresis in animal cell culture technology.
A nonrecombinant human melanoma cell line and recombinant chinese hamster ovary (CHO) cells were used as examples for long-term in vitro cultivation in protein-free media. The method used to monitor the consistency of protein release by these mammalian cells was two-dimensional electrophoresis with immobilized pH gradient. Secreted proteins from a melanoma cell line cultivated in a continuous fermentation system over a period of 22 months were monitored. Two-dimensional patterns of secreted proteins were compared and the stability of their composition was determined over a period of nearly 14 months, with significant pattern variation being observed after 14 months. The protein pattern from this extended in vitro culture was compared to those of the very same melanoma cell line recultivated after being frozen in liquid nitrogen for more than 2 years. Due to the high resolution of complex polypeptide mixtures and the possibility to detect even minor differences in the composition of protein patterns, we propose the two-dimensional electrophoresis as a tool for quality assessment in animal cell culture technology.
Institute of Applied Microbiology, University of Agriculture and Forestry, Vienna, Austria.
H
H
Harant
K
K
Wimmer
E
E
Wenisch
K
K
Strutzenberger
M
M
Reiter
G
G
Blüml
T
T
Gaida
C
C
Schmatz
H
H
Katinger
D000818:Animals; D001709:Biotechnology / Q000379:methods; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007525:Isoelectric Focusing; D011506:Proteins / Q000378:metabolism; D011786:Quality Control; D014407:Tumor Cells, Cultured
D011506:Proteins