The antigenic structure of the human glycoprotein hormone alpha-subunit. I. Characterization of anti-alpha monoclonal antibodies.
The glycoprotein hormones CG, LH, FSH, and TSH are composed of two noncovalently linked subunits, alpha and beta. The beta-subunit confers hormone specificity, while the alpha-subunit is homologous within a species. To help in determining the antigenic structure of the common alpha-subunit, six monoclonal antibodies (mAbs) to the free or heterodimeric alpha-subunit of human (h) gonadotropic hormones have been prepared and, along with two previously isolated mAbs, have been characterized for binding specificity to alpha- and beta-subunits and the human glycoprotein hormones, CG, LH, FSH, and TSH. Each mAb was derived from hybidomas of FO myeloma cells fused with spleen cells from mice immunized with free alpha-subunit, hCG or hFSH. mAbs A101, A102, and E512 were specific for the alpha-subunit but showed the highest affinity for the intact hormone; K2.18, K94.6, E501, E502, and E511 were specific for free alpha. All of the antibodies inhibited binding of 125I-hCG to luteal membrane receptor, and 125I-labeled mAbs did not recognize hCG/receptor complex. Characterization by two-site binding assays using alpha, hCG, or hFSH as antigen revealed that all the mAbs bind to unique sites on alpha which may be overlapping, and which are modified in the intact hormone. The antigenic sites for mAbs E502, E511, and K2.18 are at least partially linear because they bind to reduced, carboxymethylated alpha.
Department of Biochemistry and Molecular Biology, Mayo Medical School, Rochester, Minnesota 55905.
M C
MC
Charlesworth
D J
DJ
McCormick
E R
ER
Bergert
T
T
Vutyavanich
H
H
Hojo
R J
RJ
Ryan
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D006063:Chorionic Gonadotropin / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D005260:Female; D005640:Follicle Stimulating Hormone / Q000276:immunology; D015292:Glycoprotein Hormones, alpha Subunit / Q000032:analysis / Q000276:immunology*; D007986:Luteinizing Hormone / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D010053:Ovary / Q000378:metabolism; D011863:Radioimmunoassay; D011869:Radioligand Assay; D051381:Rats; D011967:Receptors, Gonadotropin / Q000378:metabolism; D013972:Thyrotropin / Q000276:immunology
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D000939:Epitopes; D015292:Glycoprotein Hormones, alpha Subunit; D011967:Receptors, Gonadotropin; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D013972:Thyrotropin