Cell aggregation in a Chinese hamster ovary cell microcarrier culture affects the expression rate and N-linked glycosylation of recombinant mouse placental lactogen-1.
A microcarrier culture of Chinese hamster ovary (CHO) cells, expressing the N-glycosylated recombinant protein mouse placental lactogen I (mPL-I), was found to form large cellular aggregates (400 to 600 microns in diameter). There was increased accumulation of lower molecular sized mPL-I glycoforms in cultures containing the large cellular aggregates at pH 7.3, but not at pH 7.6. Specific rates of mPL-I expression were found in the cultures with cellular aggregates at both pH values (7.3 and 7.6). These findings are interpreted in the light of our earlier studies that showed that extracellular pH and elevated ammonia concentrations affect both the glycosylation and the expression rates of mouse placental lactogen I.
Department of Chemical Engineering, Northwestern University, Evanston, Illinois 60208.
M C
MC
Borys
D I
DI
Linzer
E T
ET
Papoutsakis
D000641:Ammonia / Q000378:metabolism; D000818:Animals; D016466:CHO Cells / Q000166:cytology / Q000378:metabolism*; D002449:Cell Aggregation; D006224:Cricetinae; D003412:Cricetulus; D017077:Culture Media, Conditioned / Q000737:chemistry; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods*; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008863:Microspheres; D008970:Molecular Weight; D010928:Placental Lactogen / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000378:metabolism*
D017077:Culture Media, Conditioned; D011993:Recombinant Fusion Proteins; D000641:Ammonia; D010928:Placental Lactogen