摘要
以人工合成的降钙素基因相关肽（ＣＧＲＰ）一牛血清白蛋白（ＢＳＡ）偶联物（ＣＧＲＰ—ＢＳＡ）为免疫原，用微量脾内免疫法免疫ＢＡＬＢ／ｃ小鼠。取其免疫脾细胞与小鼠ＳＰ２／０骨髓瘤细胞融合，经间接ＥＬＩＳＡ法筛选、３次克隆化后，获得３株能稳定分泌抗ＣＧＲＰ单克隆抗体的杂交瘤细胞３Ｂ４、７Ｅ１１和８Ｆ２。用琼脂双扩散法鉴定，３Ｂ４为ＩｇＧ１亚类，７Ｅ１１和８Ｆ２均为ＩｇＭ类。杂交瘤细胞的染色体条数在９２～１１４条之间。特异性鉴定表明，３Ｂ４腹水（１：１００）与心钠素、甲状腺降钙素、绒毛膜促性腺激素和血管紧张素Ⅰ等８种抗原均无交叉反应。８Ｆ２腹水、３Ｂ４腹水及７Ｅ１１腹水的效价分别为：１．６×１０－５、３．１×１０－５及１．２５×１０－４。ＭｃＡｂ相加试验及竟争抑制试验均表明，３Ｂ４、７Ｅ１１和８Ｆ２分别识别不同的表位。以Ｗｉｓｔａｒ大白鼠为实验动物，各株杂交瘤细胞的腹水（１：１００）为样品，进行ＭｃＡｂ的动物阻断保护试验。即将大白鼠麻醉后颈动脉插管，单独注射ＣＧＲＰ（３μｇ／０．１ｍｌ只），血压下降６．６６５—９．３３３ｋＰａ；当注入ＣＧＲＰ（３μｇ）加ＭｃＡｂ８Ｆ２（１：１００）时，血压下降１１．９９７ｋｐａ，比?
An coupling complex of CGRP-BSA was used to immunize BALB/c mice via intra--spleenand three hybridoma cell lines, 3B4, 7E11 and 8F2, were obtained by fusing SpZ/0 myeloma cell lineswith the complex-primed mouse spleen cells.By double gel diffusion test, 3B4 was shown to be a lgGI,while both 7E11 and 8F2 were IgM.The number of chromosomes of hybridomas were betwen 92 to114.The McAb titers of 8F2,3B4 and 7E11 ascites is 1.5×10-5,3.1×10-5 and 1.25×10-4, respectively.Under anesthesia, the right carotid artery of experimental Wistar ratS was cannulated withPOly-ethyene tube connected to a pressure transducer for recording mean arterial pressure on aPOlygraph.When CGRP alone infuses into the rats(3ng/0.1ml each rat),the BP-decreasing value is a6.665～9.335kpa,while CGRP(3pg) plus McAb 8F2(1: 100)are administrated value of BP is 11.997kpa,2.665～5.332kpa lower than only infusion of CGRP It is suggested that binding of 8F2 to its corresponding antigen lead to futher exPOsure of CGRP's domain and so enhance the depressor resPOnse toCGRP.
