ELISA for F-TCF (human hepatocyte growth factor/hHGF)/fibroblast-derived tumor cytotoxic factor antigen employing monoclonal antibodies and its application to patients with liver diseases.
For determination of fibroblast-derived tumor cytotoxic factor, F-TCF (human hepatocyte growth factor/hHGF), sensitive two-step sandwich enzyme-linked immunosorbent assay (ELISA) employing monoclonal antibodies was developed. Microplates were coated with monoclonal antibody (P1C8) and bound F-TCF was quantitated with the second monoclonal antibody (P2D6) linked to peroxidase. The standard curve for F-TCF was found to be linear in the range of 0.16 to 10 ng of F-TCF per ml. The assay was specific for F-TCF but not for plasminogen. The assay can be used for determination of F-TCF antigen in both human plasma and serum. The variation of absorbance was little in duplicate samples. Recoveries of exogenous F-TCF added to serum or plasma samples showed theoretical values. F-TCF antigen levels in 21 healthy volunteers was found to be 0.56 +/- 0.43 ng/ml. In contrast, mean F-TCF levels in patients with liver diseases were all higher than those of healthy subjects. This ELISA system has the advantage of using a sensitive and reproducible set of monoclonal antibodies, and is a useful method for monitoring F-TCF levels in patients with liver diseases.
Shima
N
N
Life Science Research Institute, Snow Brand Milk Products Co., Ltd. Tochigi, Japan.
Higashio
K
K
Ogaki
H
H
Okabe
K
K
D000818:Animals; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006133:Growth Substances / Q000032:analysis* / Q000276:immunology; D017228:Hepatocyte Growth Factor; D006801:Humans; D008107:Liver Diseases / Q000175:diagnosis*; D008115:Liver Regeneration / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D006133:Growth Substances; D017228:Hepatocyte Growth Factor