Production of recombinant human monoclonal antibody using ras-amplified BHK-21 cells in a protein-free medium.
A ras oncogene-amplified recombinant BHK-21 cell line (ras-rBHK-IgG) has been established, and was shown to hyperproduce the recombinant IgG chimeric human monoclonal antibody (hMAb) AE6F4, which recognizes lung cancer cells. We found that the ras-rBHK-IgG cell could be easily cultured in a protein-free ERDF medium supplemented with iron(III) nitrate, hydroxyethyliminodiacetic acid, and non-protein synthetic attachment factor as well as in a serum-free ERDF medium supplemented with insulin, transferrin, ethanolamine, and sodium selenite. The productivity of recombinant hMAb from the cells cultured in dishes at high cell densities was higher in protein-free medium than in serum-containing medium. True high density culture of the ras-rBHK-IgG cells was done in protein-free medium using the Tecnomouse, which is a novel hollow fiber bioreactor system. After culture for 30 days in protein-free culture, a total amount of about 14 mg of the recombinant hMAb AE6F4 was obtained, and was shown to be reactive against lung cancer cells in tissues.
Inoue
Y
Y
Graduate School of Genetic Resources Technology, Kyushu University, Fukuoka, Japan.
López
L B
LB
Kawamoto
S
S
Arita
N
N
Teruya
K
K
Seki
K
K
Shoji
M
M
Kamei
M
M
Hashizume
S
S
Shiozawa
Y
Y
Tachibana
H
H
Ohashi
H
H
Yasumoto
K
K
Nagashima
A
A
Nakahashi
H
H
Suzuki
T
T
Imai
T
T
Nomoto
K
K
Takenoyama
M
M
Katakura
Y
Y
Shirahata
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D016466:CHO Cells; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free; D019856:Ethanolamine; D004983:Ethanolamines / Q000494:pharmacology; D005290:Ferric Compounds / Q000494:pharmacology; D006801:Humans; D007098:Imino Acids / Q000494:pharmacology; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000235:genetics; D007328:Insulin / Q000494:pharmacology; D008689:Methacrylates / Q000138:chemical synthesis / Q000494:pharmacology; D009566:Nitrates / Q000494:pharmacology; D015689:Oncogene Protein p21(ras) / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D018038:Sodium Selenite / Q000494:pharmacology; D014168:Transferrin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D016895:Culture Media, Serum-Free; D004983:Ethanolamines; D005290:Ferric Compounds; D007098:Imino Acids; D007074:Immunoglobulin G; D007328:Insulin; D008689:Methacrylates; D009566:Nitrates; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014168:Transferrin; D019856:Ethanolamine; C008021:N-(2-hydroxyethyl)iminodiacetic acid; D015689:Oncogene Protein p21(ras); D018038:Sodium Selenite; C025302:ferric nitrate