Comparison of four methods to generate immunoreactive fragments of a murine monoclonal antibody OC859 against human ovarian epithelial cancer antigen.
In the present study, four different proteases (pepsin, papain, bromelain and ficin) were screened with a murine monoclonal antibody OC859, in order to verify whether different digestion procedures could improve yield and stability of the F(ab')2 or Fab fragments. The yields of F(ab')2 or Fab fragments from digestion with pepsin, papain, bromelain and ficin were respectively 20.3 +/- 2.0%, 50.5 +/- 5.0%, 74.4 +/- 2.7% and 82.8 +/- 10.2% of the theoretical maximum. Immunoreactivity in a noncompetitive solid-phase radioimmunoassay (SPRIA) of the fragments generated by the four proteases were respectively 10 +/- 5%, 36 +/- 5%, 60 +/- 6% and 75 +/- 6% of the intact OC859 IgG. These results suggested that the fragmentation of OC859 with ficin gave a higher yield of superior immunoreactive fragments.
Zou
Y
Y
Department of Obstetrics and Gynecology, PUMC Hospital, Beijing.
Bian
M
M
Yiang
Z
Z
Lian
L
L
Liu
W
W
Xu
X
X
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000912:Antibodies, Neoplasm / Q000378:metabolism*; D000951:Antigens, Neoplasm / Q000276:immunology; D005260:Female; D005361:Ficain; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification*; D051379:Mice; D010051:Ovarian Neoplasms / Q000276:immunology; D014407:Tumor Cells, Cultured / Q000276:immunology
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D007140:Immunoglobulin Fab Fragments; D005361:Ficain