Monoclonal antibodies against human basic fibroblast growth factor.
Recombinant human basic fibroblast growth factor (hbFGF) was used as an antigen to develop, by a somatic cell fusion technique, four monoclonal antibodies (MAbs), that recognize the complete and amino-terminal truncated form of hbFGF. Isotype identification showed that MAbs designated MAb12 and MAb98 were IgG1; and those designated MAb52 and MAb78 were IgG2b. All these MAbs bound the complete form of hbFGF produced in E.coli. Competition with synthetic polypeptides, a replication of 1-9 aa and of 141-146 aa of hbFGF, and truncated forms of hbFGF by 13 and 40 amino acid residues in its amino-terminal produced in E. coli by recombinant technique, revealed at least two epitopes recognized by the four IgG type MAbs. MAb12 and MAb78 recognized the epitope located within the first 9 amino acid residues at the amino terminal of the complete hbFGF. MAb52 and MAb98 recognized the one located between the amino acid residue no. 14 and 40. None of MAbs bound bovine acidic FGF (aFGF). Using MAb52 or MAb98 and MAb78, a two-site EIA has been developed. This EIA is sensitive enough to detect 0.5 ng/ml of hbFGF. Furthermore, MAb78 was used as a ligand for affinity chromatography to purify hbFGF mutein CS4, which binds weakly to a heparin affinity column.
Seno
M
M
Biotechnology Laboratories, Takeda Chemical Ind., Ltd., Osaka, Japan.
Iwane
M
M
Sasada
R
R
Moriya
N
N
Kurokawa
T
T
Igarashi
K
K
D000818:Animals; D000911:Antibodies, Monoclonal; D003001:Cloning, Molecular; D000939:Epitopes; D005346:Fibroblast Growth Factors / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007124:Immunoenzyme Techniques; D051379:Mice
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005346:Fibroblast Growth Factors