A Monoclonal Antibody to α‐Human Atrial Natriuretic Polypeptide
A Monoclonal Antibody to α‐Human Atrial Natriuretic Polypeptide
复制标题
α-人心房钠尿多肽单克隆抗体
DOI：
10.1161/01.hyp.12.2.117
复制
发表时间：
1988
期刊：
影响因子：
8.3
通讯作者：
H. Imura
中科院分区：
文献类型：
--
作者：
M. Mukoyama;H. Sugawa;N. Morii;A. Sugawara;Takayuki Yamada;H. Itoh;S. Shiono;Yoshihiko Saito;H. Arai;Tohru Mori;H. Yamada;Y. Sano;H. Imura
用碳二亚胺偶联法，将未产生α-人心钠素（α-hANP）的小鼠骨髓瘤细胞株X63-Ag 8.653与合成的α-hANP-牛甲状腺素免疫的BALB/c小鼠脾细胞融合，制备了抗α-hANP的单克隆抗体KY-ANP-I。用125 I-α-hANP和培养液对杂交瘤进行抗体筛选。通过11 mtHng稀释技术克隆它们，在培养物中扩增，并将mtraperHoneaDy注射到BALB/c小鼠中。所获得的抗体属于免疫球蛋白G1亚类。Scatchard图分析显示对a-hANP的高亲和力，缔合常数为3.1 x 1010 M−1。用此单克隆抗体建立了特异性的α-hANP放射免疫分析法。小鼠腹水中的抗体可用于放射免疫测定，最终稀释度为1：106。本放射免疫测定法的IC_（10）和IC_（50）值分别为3和30 fmol/管。放射免疫法与α-大鼠心钠素的交叉反应率为0.9%。α-hANP-（8-22）和α-ANP-（1 -6）的交叉反应性低于α-大鼠ANP。与α-ANP-（17-28）无交叉反应。因此，识别的表位必须位于包括Met 12残基的α-hANP的环结构的N-末端的一半。该放射免疫分析法可同时检测γ-hANP、β-hANP和α-hANP。该单克隆抗体也可用于免疫组化研究。应用抗生物素蛋白-btotin-过氧化物酶复合物技术，在人心房肌中，ANP阳性细胞被精细染色。这些结果表明，抗α-hANP单克隆抗体将成为研究ANP生理和病理生理意义的有力工具。
A monoclonal antibody to α-human atrial natriuretic polypeptide (α-hANP), KY-ANP-I, has been produced by fusion of a nonproducmg mouse myeloma cell tine, X63-Ag8.653, with spleen ceDs from BALB/c mice immunized with synthetic α-hANP conjugated to bovine thyrogtobuhn using the carbodiimide coupling procedure. Hybrklomas were screened for antibody production by rarJiolmmunoassay using culture media and 125I-α-hANP. They were cloned by the llmtHng dilution technique, expanded hi culture, and injected mtraperHoneaDy into BALB/c mice. The obtained antibody belonged to the immunogiobunn G1 subclass. Analysis by a Scatchard plot revealed a high affinity for a-hANP, with an association constant of 3.1 x 1010 M−1. With this monoclonal antibody, a specific radioimmunoassay for α-hANP has been established. The antibody in mouse ascites was available for radioimmunoassay at a final dilution of 1:106. Values of IC10 and IC50 in this radioimmunoassay were 3 and 30 fmol/tube, respectively. The radioimmunoassay showed a cross-reactivity of 0.9% with α-rat ANP. α-hANP-(8–22) and α-ANP-(l–6) exhibited less cross-reactivity than α-rat ANP on a molar basis. There was no cross-reaction with α-ANP-(17–28). Thus, the recognized epitope must be located in the N-terminal half of the ring structure of α-hANP including Met12 residue. This radioimmunoassay could detect γ-hANP and β-hANP as well as α-hANP. The monoclonal antibody was also useful for unmunohistochemical studies. ANP-positive cells were finely stained hi the human atrium using the avidin-btotin-peroxidase complex technique. These results indicate that this monoclonal antibody to α-hANP will become a powerful tool for investigating the physiological and pathophysiological significance of ANP.
具有皮摩尔抗原的下丘脑生长激素释放因子单克隆抗体。
DOI：
10.1126/science.6813967
发表时间：
1982
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Luben,RA;Brazeau,P;Böhlen,P;Guillemin,R
通讯作者：
Guillemin,R
使用针对绵羊材料的单克隆抗体测定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的水平。
影响因子：
4.8
作者：
Hollander,CS;Audhya,T;Russo,M;Passarelli,J;Nakane,T;Schlesinger,D
通讯作者：
Schlesinger,D