用合成多肽免疫制备抗人心肌肌钙蛋白Ⅰ单克隆抗体的研究  

Making of monoclonal antibodiesspecific against human cardiac troponin I by synthetic multiple antigen peptide

作　　者：丛进阳[1] 盖晓东[1] 沈忠耀[1] 

机构地区：[1]清华大学化学工程系

出　　处：《清华大学学报（自然科学版）》1997年第12期55-59,共5页Journal of Tsinghua University(Science and Technology)

基　　金：国家自然科学基金

摘　　要：介绍一种用合成的多抗原肽免疫动物来制备与天然人心肌肌钙蛋白Ⅰ特异反应的单克隆抗体新方法。从人心肌肌钙蛋白ⅠＮ端，根据多肽分子氨基酸的亲水性值选出了ＰＡＰＡＰＩＡＡＡＳＳ１１个氨基酸片段。将它用有八位点的赖氨酸核连接成多抗原肽后，以它为抗原用Ｒｉｂｉ佐剂进行免疫。将小鼠脾细胞与ＳＰ２／０骨髓瘤细胞融合，获１１株抗此多肽的杂交瘤细胞株。对其中７株属于ＩｇＧ亚类的细胞进行了ＥＬＩＳＡ分析，发现这７株细胞均与天然的人心肌肌钙蛋白Ⅰ呈特异性反应，而不与人骨骼肌肌钙蛋白Ⅰ发生交叉反应。利用３Ａ２细胞株所分泌的单克隆抗体对急性心肌梗塞患者、手术患者和正常人的血样进行了ＥＬＩＳＡ检测，证明它只对心肌梗塞患者血清发生阳性反应。获得了较为满意的结果。此细胞株所分泌的抗体有望用于心肌梗塞临床检测。A new method for making hybridoma cell lines producing monoclonal antibodies specific for human cardiac troponin I by immunization of animals with synthetic peptides has been introduced. Eleven amino acid sequence PAPAPIAAASS was selected from the Nterminus of human cardiac troponin Ⅰ, and linked to lysine core with 8branching sites to form a multiple antigen peptide (MAP). The MAP was used as an antigen, and mixed with Ribiadjuvant system for immunizing BALB/c mice. Fusions of spleen cells from the mice with SP2/0 myeloma cells were made, and 11 positive hybridoma cell lines against the MAP were obtained after fusions. Among them 7 cell lines belong to IgG subclasses. In further investigation of these 7 cell lines by ELISA, it was indicated that all the 7 cell lines had specific reactions with native human cardiac troponin Ⅰ, and did not react with human skeletal troponin Ⅰ. The sera of patients suffering from acute myocardial infarction, patients subjected to operations, and normal health persons were tested by ELISA with monoclonal antibody produced by 3A2 cell line. The results show that only patients with acute myocardial infarction give positive reactions in ELISA.

关 键 词：心肌梗塞 单克隆抗体 肌钙蛋白 ELISA检测 

分 类 号：R542.220.4[医药卫生—心血管疾病]