Abstract
Two mouse myeloma cell lines which were transfected with chimeric mouse variable-human constant immunoglobulin heavy and light chain genes have been cultured at high cell density in a settling perfusion culture vessel to produce chimeric antibody specific for human common acute lymphocytic leukemia antigen (cALLA).
J558L transfectant proliferated well in a serum-free medium (ITES-eRDF) to a viable cell density of 3.7×107 cells/ml and produced chimeric antibody to a maximum value of 60 μg/ml in 120 ml scale vessel. X63Ag8.653 transfectant reached a density of 1.9×107 cells/ml in 1.2 I scale vessel in serum supplemented medium (10% FCS-eRDF) and produced chimeric antibody which consisted of chimeric gamma and chimeric kappa chains to a maximum value of 5.8 μg/ml.
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References
BetterM, ChangCP, RobinsonRR and HorwitzAH (1988) Escherichia coli secretion of an active chimeric antibody fragment. Science 240: 1041–1043.
BossMA, KentenJH, WoodCR and EmtageJ (1984) Assembly of functional antibodies from immunoglobulin heavy and light chains synthesized in E. coli. Nucleic Acids Res. 12: 3791–3806.
BoulianneGL, HozumiN and ShulmanMJ (1984) Production of functional chimeric mouse/human antibody. Nature 312: 643–646.
CabillyS, RiggsAD, PandeH, ShivelyJE, HolmesWE, ReyM, PerryLJ, WetzelR and HeynekerHL (1984) Generation of antibody activity from immunoglobulin poly-peptide chains produced in Escherichia coli. Proc. Natl. Acad. Sci. USA 81: 3273–3277.
DuhamelRC, SchurPH, BrendelK and MeezanE (1979) pH gradient elution of human IgG1, IgG2 and IgG2 and IgG4 from protein A-Sepharose. J. Immunol. Methods 31: 211–217.
FujinagaS, SuganoT, MatsumotoY, MasuhoY and MoriR (1987) Antiviral activities of human monoclonal antibodies to herpes simplex virus. J. Infect. Dis. 155: 45–53.
LiuAY, MackPW, ChampionCI and RobinsonRR (1987a) Expression of mouse: human immunoglobulin heavy-chain cDNA in lymphoid cells. Gene 54: 33–40.
LiuAY, RobinsonRR, HellstromKE, MurrayJrED, ChangCP and HellstromI (1987b) Chimeric mouse-human IgG1 antibody that can mediate lysis of cancer cells. Proc. Natl. Acad. Sci. USA 84: 3439–3443.
LiuAY, RobinsonRR, MurrayJrED, LedbetterJA, HellstromI and HellstromKE (1987c) Production of a mouse-human chimeric monoclonal antibody to CD20 with potent Fc dependent biologic activity. J. Immunol. 139: 3521–3526.
MorrisonSL, JohnsonMJ, HerzenbergLA and OiVT (1984) Chimeric human antibody molecules: Mouse antigen-binding domains with human constant region domains. Proc. Natl. Acad. Sci. USA 81: 6851–6855.
MurakamiH, MasuiH, SatoGH, SueokaN, ChowTP and Kano-SueokaT (1982) Growth of hybridoma cells in serum-free medium. Proc. Natl. Acad. Sci. USA 79: 1158–1162.
MurakamiH, ShimomuraT, NakamuraT, OhashiH, ShinoharaK and OmuraH (1984) Development of a basal medium for serum-free cultivation of hybridoma cells in high density. Nippon Nogeikagaku Kaishi 58: 575–583.
NeubergerMS, WilliamsGT and FoxRO (1984) Recombinant antibodies processing novel effector functions. Nature 312: 604–608.
NeubergerMS, WilliamsGT, MitchellEB, JouhalSS, FlanaganJG and RobbittsTH (1985) A hapten-specific chimeric IgE antibody with human physiological effector function. Nature 314: 268–270.
NishimuraY, YokoyamaM, ArakiK, UedoR, KudoA and WatanbeT (1987) Recombinant human-mouse chimeric monoclonal antibody specific for common acute lymphocytic leukemia antigen. Cancer research 47: 999–1005.
OiVT, MorrisonSL, HerzenbergLA and BergP (1983) Immunoglobulin gene expression in transformed lymphoid cells. Proc. Natl. Acad. Sci. USA 80: 825–829.
RhodesM and BirchJ (1988) Large scale production of proteins from mammalian cells. Bio/technology 6: 518–523.
SahaganBG, DoraiH, Saltzgaber-MullerJ, ToneguzzoF, GuindonCA, LillySP, McDonaldKW, MorrisseyDV, StoneBA, DavisGL, McIntoshPK and MoorGP (1986) A genetically engineered murine/human chimeric antibody retains specificity for human tumor-associated antigen. J. Immunol. 137: 1066–1074.
SunLK, CurtisP, Rakowicz-SzulczynskE, GhrayebJ, ChangN, MorrisonSL and KoprowskiH (1987) Chimeric antibody with human constant regions and mouse variable regions directed against carcinoma-associated antigen 17-1A. Proc. Natl. Acad. Sci. USA 84: 214–218.
TakazawaY, TokashikiM, HamamotoK and MurakamiH (1988) High cell density perfusion culture of hybridoma cells recycling high molecular weight components. Cytotechnology 1: 171–178.
VerhoeyenM, MilsteinC and WinterG (1988) Reshaping human antibodies: Grafting an antilysozyme activity. Science 239: 1534–1536.
YokoyamaM, NishimuraY and WatanabeT (1987) Suppression of tumor growth by in vivo administration of recombinant human-mouse chimeric monoclonal antibody. Jpn. J. Cancer Res. (Gann) 78: 1251–1257.
Zemel-DreasenO and ZamirA (1984) Secretion and processing of an immunoglobulin light chain in Escherichia coli. Gene 27: 315–322.
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Takazawa, Y., Tokashiki, M. Production of human-mouse chimeric antibody by high cell density perfusion culture. Cytotechnology 2, 95–101 (1989). https://doi.org/10.1007/BF00386141
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DOI: https://doi.org/10.1007/BF00386141