Immunobiological properties of a carboxy-terminal 53-amino acid peptide of the beta subunit of human chorionic gonadotropin.
Lengthening of the carboxy terminus unique region of beta-hCG from 30 to 45 amino acids was found in previous studies to improve immunogenicity and hormone neutralization capacity. The present study was carried out to determine whether further elongation of the peptide to 53 amino acids enhances to hormone neutralization capacity without loss of specificity characteristics. The peptide 93--145 of beta-hCG with substitution of cysteines at 93, 100 and 110 by alpha-aminobutyric acid was synthesized by solid phase and conjugated to tetanus toxoid by an active ester method. Rabbit antibodies against this conjugate reacted with CTP-53 and CTP-45 with a parallel slope in anti-CTP-53-[125I]Tyr-CTP--53 radioimmunoassay system. Other CTPs, e.g. CTP-26, CTP-31 and CTP-35 competed with lower efficiency; 50% inhibition of binding was obtained with 10-100 pmol/tube with these peptides instead of 0.5 pmol/tube for the homologous CTP-53. Anti-CTP-53 reacted with both beta-hCG and hCG but around twenty times greater amount of hCG was required to give 50% inhibition of binding as compared to CTP-53. The antigen binding capacity of anti-CTP-53 was around 4000 ng/ml for CTP-53 and 25 ng/ml for hCG. The anti-CTP-53 sera retained non-cross-reactivity with hLH as determined by direct binding with [125I]hLH and by competitive inhibition of CTP-53 binding with anti-CTP-53. Anti-CTP-53 neutralized the bioactivity of hCG in the Leydig cell bioassay and in the mouse uterine weight gain assay. Anti-CTP-53 antibodies were about three times more effective than anti-CTP-45 in their capacity to neutralize the bioactivity of hCG, though still substantially poorer than anti-beta-hCG sera in this respect.
D000596:Amino Acids / Q000276:immunology*; D000818:Animals; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002096:C-Peptide / Q000276:immunology; D006063:Chorionic Gonadotropin / Q000276:immunology* / Q000378:metabolism; D018997:Chorionic Gonadotropin, beta Subunit, Human; D003429:Cross Reactions; D005260:Female; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D007985:Leydig Cells / Q000378:metabolism; D007986:Luteinizing Hormone / Q000378:metabolism; D008297:Male; D051379:Mice; D009929:Organ Size; D010446:Peptide Fragments / Q000276:immunology*; D011247:Pregnancy; D011817:Rabbits; D013739:Testosterone / Q000096:biosynthesis; D014599:Uterus / Q000033:anatomy & histology
D000596:Amino Acids; D002096:C-Peptide; D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D007106:Immune Sera; D010446:Peptide Fragments; D013739:Testosterone; D007986:Luteinizing Hormone