Purification of bispecific F(ab')2 from murine trinoma OC/TR with specificity for CD3 and ovarian cancer.
A study was made of the stability of the murine bispecific trinoma OC/TR with respect to secretion of both types of parental heavy and light chains. OC/TR is a cell line producing bispecific antibody that reacts with the CD3 antigen on T cells and the folate-binding receptor--frequently found to be overexpressed on ovarian carcinoma cells. Of the 10 different IgG combinations theoretically possible with 2 heavy and 2 light chains, 6 combinations were secreted. Subclones varied considerably in relative production of the two parental heavy and light chains. A detailed analysis was made of the binding characteristics and retargeting activity of each of the IgGs produced. From a clone producing a relatively high quantity of bispecific IgG, a large-scale production was initiated. The purification of clinical grade bispecific F(ab')2 from harvest fluids is described. The yield from this purification process was found to be comparable to the yield of bispecific F(ab')2 after chemical cross-linking of two different Fab'.
Warnaar
S O
SO
Centocor B.V., Leiden, The Netherlands.
De Paus
V
V
Lardenoije
R
R
Machielse
B N
BN
De Graaf
J
J
Bregonje
M
M
Van Haarlem
H
H
D000818:Animals; D018033:Antibodies, Bispecific / Q000276:immunology* / Q000302:isolation & purification; D000912:Antibodies, Neoplasm / Q000276:immunology* / Q000302:isolation & purification; D017252:CD3 Complex / Q000276:immunology*; D002352:Carrier Proteins / Q000276:immunology*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D058973:Folate Receptors, GPI-Anchored; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007140:Immunoglobulin Fab Fragments / Q000276:immunology* / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000276:immunology; D051379:Mice; D009363:Neoplasm Proteins / Q000276:immunology*; D010051:Ovarian Neoplasms / Q000276:immunology*; D011956:Receptors, Cell Surface; D013601:T-Lymphocytes / Q000276:immunology
D018033:Antibodies, Bispecific; D000912:Antibodies, Neoplasm; D017252:CD3 Complex; D002352:Carrier Proteins; D058973:Folate Receptors, GPI-Anchored; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D009363:Neoplasm Proteins; D011956:Receptors, Cell Surface