Bidart J M, Ozturk M, Bellet D H, Jolivet M, Gras-Masse H, Troalen F, Bohuon C J, Wands J R
J Immunol. 1985 Jan;134(1):457-64.
We have produced a library of monoclonal antibodies directed against a 37-amino acid synthetic polypeptide analogous to the carboxyl terminus of hCG. Five antibodies, designated FB01, FB02, FB03, FB04, and FB00, were developed and analyzed with respect to affinity and specificity for epitopes on human chorionic gonadotropin (hCG) and beta hCG by enzyme-linked immunoabsorbent and radioimmunoassays (RIA). All monoclonal antibodies demonstrated low affinity constants (approximately 10(-7) liters/mol) compared with those obtained by immunization with native beta hCG. One antibody, namely FB00, bound only to the synthetic peptide, whereas all other monoclonal antibodies recognized either free native beta hCG or both beta hCG and HCG. Antibodies produced against the synthetic peptide did not cross-react with other glycoprotein hormones such as LH, TSH, and FSH. Characterization of the monoclonal antibody-binding sites revealed the presence of at least four separate and distinct epitopes on the last 35 amino acids of beta hCG. Indeed, one epitope recognized by FB01 is located between residues 109 and 118, whereas another antigenic region recognized by FB04 appears to be present on the 109-121 portion of the molecule near or at position 118. One additional antigenic site was localized between residues 118 and 136. Finally, FB00 recognized an epitope located on the last 10 amino acids (136-145) of beta hCG. With the use of such antibodies, two- and three-site monoclonal RIA were developed and employed to detect free beta hCG and hCG in sera of patients with choriocarcinoma. These assays may be useful in the detection of beta hCG- and hCG-producing tumors and subsequent monitoring of patients in response to surgery and/or chemotherapy.
我们制备了一个单克隆抗体文库，这些抗体针对一种37个氨基酸的合成多肽，该多肽类似于人绒毛膜促性腺激素（hCG）的羧基末端。通过酶联免疫吸附测定和放射免疫测定（RIA），开发并分析了五种名为FB01、FB02、FB03、FB04和FB00的抗体，以检测它们对人绒毛膜促性腺激素（hCG）和β-hCG表位的亲和力和特异性。与用天然β-hCG免疫获得的抗体相比，所有单克隆抗体的亲和常数都较低（约10^(-7)升/摩尔）。一种名为FB00的抗体仅与合成肽结合，而所有其他单克隆抗体则识别游离的天然β-hCG或同时识别β-hCG和hCG。针对合成肽产生的抗体不会与其他糖蛋白激素如促黄体生成素（LH）、促甲状腺激素（TSH）和促卵泡激素（FSH）发生交叉反应。单克隆抗体结合位点的表征显示，在β-hCG的最后35个氨基酸上存在至少四个不同的表位。实际上，FB01识别的一个表位位于第109至118位氨基酸之间，而FB04识别的另一个抗原区域似乎存在于分子的109 - 121部分，靠近或位于第118位氨基酸处。另一个抗原位点位于第118至136位氨基酸之间。最后，FB00识别位于β-hCG最后10个氨基酸（136 - 145）上的一个表位。利用这些抗体，开发了双位点和三位点单克隆RIA，并用于检测绒毛膜癌患者血清中的游离β-hCG和hCG。这些检测方法可能有助于检测产生β-hCG和hCG的肿瘤，并在后续监测患者对手术和/或化疗的反应。
