Hashimura E, Shimizu F, Nishino T, Imagawa K, Tateishi K, Hamaoka T
J Immunol Methods. 1982 Dec 30;55(3):375-87. doi: 10.1016/0022-1759(82)90097-7.
Antibody specific for the amino-terminal region of cholecystokinin octapeptide (CCK-8) was generated in a highly reproducible way in New Zealand white rabbits by a novel immunization procedure which involves immunization with CCK-8 peptide conjugate coupled with keyhole limpet hemocyanin (KLH) and inhibiting cross-reacting antibody formation by treatment of the animals with a potent tolerogenic conjugate of beta-alanyl-tetragastrin and a copolymer of D-glutamic acid and D-lysine (D-GL). The antisera thus produced specifically react with an amino-terminal region of CCK-8 but not with the non-sulfate form of CCK-8, nor with the carboxy-terminal region which shares a cross-reactive determinant among gastrin and cholecystokinin-related peptides (caerulein, CCK-4, CCK-8, CCK-33 and CCK-39). The antisera produced by this method allowed us to measure specifically CCK in extracts from tissue such as duodenum containing gastrin and CCK at comparable levels.
通过一种新型免疫程序，在新西兰白兔中以高度可重复的方式产生了针对胆囊收缩素八肽（CCK - 8）氨基末端区域的抗体。该免疫程序包括用与钥孔血蓝蛋白（KLH）偶联的CCK - 8肽缀合物进行免疫，并通过用β - 丙氨酰 - 四肽胃泌素与D - 谷氨酸和D - 赖氨酸的共聚物（D - GL）处理动物来抑制交叉反应性抗体的形成。由此产生的抗血清特异性地与CCK - 8的氨基末端区域反应，但不与CCK - 8的非硫酸化形式反应，也不与在胃泌素和胆囊收缩素相关肽（蛙皮素、CCK - 4、CCK - 8、CCK - 33和CCK - 39）之间共享交叉反应决定簇的羧基末端区域反应。通过这种方法产生的抗血清使我们能够特异性地测量来自含有胃泌素和CCK且水平相当的组织（如十二指肠）提取物中的CCK。
