Effect of culture conditions on IgM antibody structure, pharmacokinetics and activity.
Culture conditions affect the binding activity, charge heterogeneity, conformational stability, glycosylation, and pharmacokinetics of human monoclonal IgM HMAB-10058. The 10058 human/human/murine trioma was grown in serum-free airlift suspension culture, hollow fiber perfusion culture, or in nude mouse ascites. The ascites-produced antibody showed reduced conformational stability, greater charge and glycoform heterogeneity, and a lower average degree of sialylation than the in vitro culture-produced material. Mean residence time after IV injection in rats was approximately 80-fold greater for the ascites culture-produced material, but specific binding activity was less than 5% of that for the airlift-produced material. In vitro culture in serum-supplemented media (in a hollow fiber perfusion reactor or in shake-flasks) resulted in antibody with pharmacokinetics intermediate between the serum-free airlift and ascites-produced materials. Incubation of airlift-produced antibody in ascites fluid also resulted in material with intermediate pharmacokinetics. Conclusions regarding the effect of culture conditions on antibody product cannot be generalized, as in vitro-produced antibody derived from two related cell lines (HMAB-10233 and HMAB-10390) had long mean residence times similar to that of ascites-produced HMAB-10058.
Maiorella
B L
BL
Chiron Corp., Emeryville, CA 94608.
Winkelhake
J
J
Young
J
J
Moyer
B
B
Bauer
R
R
Hora
M
M
Andya
J
J
Thomson
J
J
Patel
T
T
Parekh
R
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D001201:Ascites; D001769:Blood; D002460:Cell Line; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004563:Electrochemistry; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007075:Immunoglobulin M / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D019158:N-Acetylneuraminic Acid; D009844:Oligosaccharides / Q000378:metabolism; D011487:Protein Conformation; D012794:Sialic Acids / Q000378:metabolism; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M; D009844:Oligosaccharides; D012794:Sialic Acids; D019158:N-Acetylneuraminic Acid