Identification of assembled epitopes on the alpha-subunit of human follicle stimulating hormone.
In order to characterize further the antigenic structure of human follitropin (hFSH), BALB/c mice were immunized with hFSH and anti-hFSH monoclonal antibodies (mAbs) were generated. The hFSH subunit specificity of the mAbs was assessed by a solid-phase enzyme-linked immunosorbent assay (ELISA) and a solution-phase radioimmunoassay (RIA), each using hFSH, hFSH alpha, and hFSH beta. Five mAbs bound hFSH and hFSH alpha in the ELISA and the RIA. In addition, some mAbs recognized hFSH beta, albeit to a much lower degree, as demonstrated by displacement of [125I]hFSH binding to the mAbs by hFSH beta, in the solution-phase RIAs. Next, synthetic peptides corresponding to the hFSH alpha-subunit sequence were used to identify sequences specific to the epitopes of each of the five mAbs. Using this epitope mapping strategy, two assembled epitopes were identified. mAbs 3A and 4B distinguish one discontinuous epitope comprised minimally of sequences alpha-16-21 and alpha-66-92, whereas mAbs 5F and 2E distinguish a second discontinuous epitope comprised minimally of sequences alpha-40-50 and alpha-66-72.
Weiner
R S
RS
Department of Biochemistry and Molecular Biology, Albany Medical College, NY 12208.
Dias
J A
JA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D005640:Follicle Stimulating Hormone / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005640:Follicle Stimulating Hormone