title: Determination of hyperglycosylated human chorionic gonadotropin produced by malignant gestational trophoblastic neoplasias and male germ cell tumors using a lectin-based immunoassay and surface plasmon resonance.
abstract: The ability to reliably detect aberrant glycosylation of human chorionic gonadotropin (hCG) may have profound implications for the diagnosis and monitoring of malignant gestational trophoblastic neoplasia, germ cell tumors, other malignancies, and pregnancy complications. To become a clinically useful assay, however, this discrimination of glycoforms should be possible on minimally treated biological specimens. Towards this end, we have developed a lectin-based sandwich-type immunoassay to compare the glycosylation patterns of hCG among urine specimens from patients presenting with a normal pregnancy, invasive mole, choriocarcinoma, and male germ cell tumors using carbohydrate-free antibody fragments as capture reagents and a panel of eight lectins, five recognizing neutral sugars and three recognizing sialic acid. There was no significant difference in the binding of any of the lectins to hCG in the urine of women over the gestational range of 6-38 weeks. Three lectins, however, exhibited differential binding to urinary hCG derived from these normal pregnant controls and that from patients with malignant forms of gestational trophoblastic disease and male germ cell tumors. Galanthus nivalis agglutinin and Maackia amurensis lectin, which bind terminal mannose and alpha(2-3)sialic acid, respectively, preferentially bound pregnancy-derived hCG, whereas the lectin, wheat germ agglutinin, which binds sialic acid and beta(1-4)N-acetylglucosamine, exhibited decreased binding to pregnancy-derived hCG compared to that from patients with male germ cell tumors and malignant gestational trophoblastic neoplasia. The differential binding observed with these three promising lectins is most encouraging and warrants further examination. The experimental paradigm also holds promise for the development of comparable assays for other glycosylated tumor markers.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, United States.
authors.forename: Lisa S
authors.initials: LS
authors.lastname: Kelly
authors.forename: Steven
authors.initials: S
authors.lastname: Birken
authors.forename: David
authors.initials: D
authors.lastname: Puett
mesh_terms: D002241:Carbohydrates; D006063:Chorionic Gonadotropin / Q000652:urine*; D005260:Female; D031901:Gestational Trophoblastic Disease / Q000473:pathology* / Q000652:urine*; D006031:Glycosylation; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D037102:Lectins / Q000378:metabolism*; D008297:Male; D019158:N-Acetylneuraminic Acid; D009373:Neoplasms, Germ Cell and Embryonal / Q000473:pathology* / Q000652:urine*; D011247:Pregnancy; D011485:Protein Binding; D020349:Surface Plasmon Resonance
chemical_list: D002241:Carbohydrates; D006063:Chorionic Gonadotropin; D037102:Lectins; C024889:glycosylated HCG; D019158:N-Acetylneuraminic Acid