摘要
利用基因重组技术，大肠杆菌表达人环胞菌素Ａ结合蛋白（ｒｈＣｙＰＡ），建立测定抗ＣｙＰＡ自身抗体的酶联免疫吸附试验（ＥＬＩＳＡ），探讨抗ＣｙＰＡ抗体与临床疾病的关系。重组菌可溶性蛋白经饱和硫酸铵盐析和ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ柱层析，获取ｒｈＣｙＰＡ蛋白，用该纯化物作为抗原，方阵滴定，建立ＥＬＩＳＡ技术，对各类自身免疫病进行评估测定。ｒｈＣｙＰＡ经十二烷基硫酸钠－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）分析，分子量为１８ｋＤ。在系统性红斑狼疮（ＳＬＥ）患者中，发现有４０．３％的病人检出有抗ＣｙＰＡ自身抗体（５８／１４４），三类Ｉｇ中，又以ＩｇＧ类抗体检出率最为突出，为２８％（４０／１４４）；而其他自身免疫病病人抗ＣｙＰＡ抗体总体检出率为０％～１８．１％。
Human cyclophilin A(CyP A)was expressed in Escherichia coli by gene recombination technique,developing enzyme linked immunosorbent assay(ELISA)for CyP A autoantibodies.The aim of prescent study was to investigate the relationship between these antibodies and clinical diseases.Saturated(NH 4) 2SO 4 salting out the dissoluble proteins of recombinant germline,and passed through a DEAE Sepharose CL 6 B column,followed by obtaining rhCyP A protein.Using the purification as antigen,square titration,and ELISA for antibodies to CyP A was developed,and was used to evaluate various autoimmune diseases.rhCyP A was analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS PAGE),and its molecular weight(MW)was 18 kD. Anti CyP A autoantibodies were found in the sera of 40 3%(58/144)of patients with systemic lupus erythematosus(SLE),IgG class antibody was most prevelance in three classes of immunoglobulin(Ig),and its possitive rate was 27 8%(40/144);while,compared with 0%～18.1% of patients with other autoimmune diseases.Suggesting that the presence of anti CyP A autoantibodies appears to be associated withSLE.
出处
《临床检验杂志》
CAS
CSCD
北大核心
1998年第5期262-265,共4页
Chinese Journal of Clinical Laboratory Science
