Enzyme Immunoassay of Urinary -Core Fragment of Human Chorionic Gonadotropin as a Tumor Marker for O
Human chorionic gonadotropin (hCG), a glycoprotein hormone composed of two nonidentical α- and β-subunits, is normally produced by trophoblasts. Serum and urine from some patients with nontrophoblastic tumors are found to contain similar immunoactivity to the β-subunit of hCG and this phenomenon has been recognized as an ectopic hCGβ production ( 1 ). Elevated levels of ectopic hCGβ are detected more frequently in urine than in serum. Recent studies have shown that urinary ectopic hCGβ mainly represents hCGβ-core fragment (β-CF) ( 2 ). Urinary β-CF consists of residues 6-40 disulfide bridged to residues 55-92 of the β-subunit of hCG ( 3 , 4 ), suggesting that it may be a degradative product of hCGβ. Such partial identity of the amino acid sequence between intact hCG, free hCGβ, and β-CF makes their specific measurements difficult. The existence of two more antigenic sites unique to hCGβ has been demonstrated. The one domain locates in the β-core portion and the other in COOH- terminal region of the β-subunit of hCG Fig. 1 ). By selecting appropriate pairs of antibodies to construct sandwich enzyme immunoassays (EIAs), it is possible to design methods that measure either intact hCG, free hCGβ, or β-CF Table 1 ). By using this EIA, the authors assessed the clinical usefulness of urinary β-CF as a tumor marker for nontrophoblastic tumors ( 5 , 6 ). Here, the authors describe the methods to measure urinary β-CF in patients with ovarian cancer. Fig. 1. Two antigenic sites: the first domain is located in the β-core portion and the second domain is located in the COOH-terminal region of the β-subunit of hCG. Table 1 Specificities and Sensitivities of Enzyme-Immunoassays Specific for Intact hCG(EIA-1), Free hCGβ(EIA-2), and β-CF(EIA-3)
First antibody | Second antibody | Cross-reactivity (%) | |||||||
---|---|---|---|---|---|---|---|---|---|
Sensitivity | Intact | Free | |||||||
Assay | Code | Epitope | Code | Epitope | Specificity | (ng/ml) | hCG | hCGβ | β-CF |
EIA-1 | No.277(MoAb) | β-CTP | No.224(MoAb) | hcGa | Intact hCG | 0.01 | 100 | 15 | 1.0 |
EIA-2 | No.209(MoAb) | hCGβ | No.115(PoAb) | β-core | Free hCGβ | 0.01 | 1.9 | 100 | 1.9 |
EIA-3 | No.229(MoAb) | β-core | No.105(PoAb) | β-core | β-CF | 0.01 | 3.3 | 100 | 100 |
MoAb: monoclonal antibody. PoAb: polyclonal antibody.