Routine large-scale production of monoclonal antibodies in a protein-free culture medium

Cleveland, William L.; Wood, Irene; Erlanger, Bernard F.

doi:10.1016/0022-1759(83)90414-3

Cited by 97 publications

(15 citation statements)

References 23 publications

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“…However, autooxidation of cholesterol occurs during handling of culture media and oxysterols formed inhibit sterol synthesis and cell (Koldovsky and Thornburg, 1987). Progesterone has sometimes been added to defined culture media for hybridoma proliferation (Cleveland et al, 1983).…”

Section: Resultsmentioning

confidence: 99%

Biochemical aspects of a whey fraction capable of promoting hybridoma proliferation. Comparison with fetal calf serum

Derouiche

Legrand

Bour

et al. 1990

Lait

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Summary -Human and bovine milks were used as serum substitute for cell cultures. Two whey fractions termed LBS65 and LELm prepared by different steps were routinely used for culturing hybridomas in our laboratory. They stimulated 3H-thymidine incorporation into hybridoma, but a minimum of 1% of fetal calf serum (FCS) was required when the cells were propagated in the long-term. ln this investigation, the chemical composition of LBS65 and LELm was studied in parallel to FCS composition. LBS65 and FCS had a total protein level of about 40 g/l, while LELm contained them at a concentration of 11 g/l. The amino acid content of FCS and LBS65 was very similar (about 4 000 urnol/l), whereas it was lower in LELm (about 300 umorl). Although the values of nitrogen cornpounds were different in LBS65 and LELm, these 2 whey fractions behaved similarly to a substitute of FCS. Whey fractions were low in sodium, and high in calcium compared to FCS. The cholesterol level was lower and triglycerides were higher in whey than in FCS. The ratio of unsaturated! saturated fatty acids varied from 1 whey fraction to another. It differed from the FCS ratio. FCS and whey fractions contained small amounts of steroid or peptide hormones. However, progesterone, 17-0H-progesterone, ACTH and cAMP concentrations were higher in the whey fraction than in FCS. Whey fractions are a complex medium containing various compounds able to promote cell growth. However, sorne unknown compounds present in FCS and absent in milk fractions are required in long-term cell culturing.bovine milk 1 whey 1 whey hormone 1 whey amino acid

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Section: Resultsmentioning

confidence: 99%

Biochemical aspects of a whey fraction capable of promoting hybridoma proliferation. Comparison with fetal calf serum

Derouiche

Legrand

Bour

et al. 1990

Lait

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“…The employment of culture vessels larger than 1 liter required addition of 0.5 ppm of antifoam A (Sigma). The large numbers of L-33 ϩ cells were then pelleted at 500 ϫ g for 5 min and resuspended at about 1 ϫ 10 6 cells/ml in three vessels each containing 16 liters of serum-free medium consisting of SMEM with 1% Nutridoma SP (Boehringer Mannheim) (v/v), 1 g/liter pluronic F68, 2 mM L-glutamine (Life Technologies, Inc.), 100 g/ml streptomycin, 25 mM HEPES, pH 8.0, 0.8 mM Na 2 SO 4 , 10 ng/ml biotin (Pierce), 50 M CaCl 2 , 0.5 ppm antifoam A, and 1 ϫ Trace Metal Mix based on previously published formulations (26,27 3 . The large suspension culture volumes employed required sparging (4 -8-m pore size air dispersion tube) (Ace Glass) with 5% CO 2 in air at a rate of 2 ml/min to maintain viability.…”

Section: Cell Culture and Collection Of Serum-free Mediummentioning

confidence: 99%

Purification of Heparan Sulfate D-Glucosaminyl 3-O-Sulfotransferase

Liu

Shworak

Fritze

et al. 1996

Journal of Biological Chemistry

118

159

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“…CCL-77). All cells were grown at 37.0°C in BM+1/2 TE1+TE2 +10% fetal calf serum (FCS) [19]. For microscope observation, cells in culture medium were dispensed into polystyrene 96-well microplates, which have glass bottoms that are 0.175 mm thick.…”

Section: Materials and Experimental Conditionsmentioning

confidence: 99%

Multiclass cell detection in bright field images of cell mixtures with ECOC probability estimation

Long

Cleveland

Yao

2008

Image and Vision Computing

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Routine large-scale production of monoclonal antibodies in a protein-free culture medium

Cited by 97 publications

References 23 publications

Biochemical aspects of a whey fraction capable of promoting hybridoma proliferation. Comparison with fetal calf serum

Biochemical aspects of a whey fraction capable of promoting hybridoma proliferation. Comparison with fetal calf serum

Purification of Heparan Sulfate D-Glucosaminyl 3-O-Sulfotransferase

Multiclass cell detection in bright field images of cell mixtures with ECOC probability estimation

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