用针对羊源物质的单克隆抗体测定的哺乳动物下丘脑和下丘脑外脑组织中促肾上腺皮质激素释放因子样免疫反应性水平。

Levels of corticotropin releasing factor-like immunoreactivity in mammalian hypothalamic and extrahypothalamic brain tissue as determined with a monoclonal antibody to the ovine material.

作者信息

Hollander C S, Audhya T, Russo M, Passarelli J, Nakane T, Schlesinger D

出版信息

Endocrinology. 1983 Jun;112(6):2206-8. doi: 10.1210/endo-112-6-2206.

PMID:6602048

Abstract

A monoclonal antibody to ovine corticotropin releasing factor (CRF) has been produced by fusion of a non-producing plasmacytoma cell line P3U1 with spleen cells of Balb/c mice immunized with the synthetic 41 amino acid peptide coupled covalently with rabbit myosin by a heterobifunctional reagent, N-succinimidyl 3-(2-pyridyldithio) propionate. A total immunizing dose of 500 micrograms resulted in a highly specific, high-affinity antibody with a Ka of 0.15 x 10(12) M-1, which was used to establish a specific RIA with a sensitivity of 10 pg/tube. Levels of corticotropin releasing factor-like immunoreactivity (CRF-LI) in a pg/mg of hypothalamic tissue ranged from 4-10 in ovine, 2.5-8 in bovine, 47.5-67.5 in mouse and 2.3-20 in human tissue. Moreover, CRF-LI was widely distributed in extrahypothalamic mouse brain at concentrations approximately one half those seen in hypothalamus.

摘要

通过非分泌性浆细胞瘤细胞系P3U1与经合成的41个氨基酸的肽免疫的Balb/c小鼠脾细胞融合，制备了一种抗绵羊促肾上腺皮质激素释放因子（CRF）的单克隆抗体。该合成肽通过异双功能试剂N-琥珀酰亚胺基-3-（2-吡啶基二硫代）丙酸与兔肌球蛋白共价偶联。500微克的总免疫剂量产生了一种高度特异性、高亲和力的抗体，其解离常数Ka为0.15×10¹² M⁻¹，该抗体用于建立灵敏度为10 pg/管的特异性放射免疫分析（RIA）。每毫克下丘脑组织中促肾上腺皮质激素释放因子样免疫反应性（CRF-LI）的水平在绵羊中为4 - 10 pg/mg，牛中为2.5 - 8 pg/mg，小鼠中为47.5 - 67.5 pg/mg，人组织中为2.3 - 20 pg/mg。此外，CRF-LI广泛分布于小鼠下丘脑外的脑组织中，其浓度约为下丘脑浓度的一半。