特异性亲环素A结合法测定环孢素血药浓度  

Determination of cyclosporine A in blood with speific cyclophilin A binding assay

作　　者：李芳秋[1] 张中书[1] 许瑞吉[1] 储小曼[1] 周燕[1] 武建国[1] 

机构地区：[1]南京军区南京总医院全军医学检验中心,江苏南京210002

出　　处：《中国药学杂志》2000年第8期536-538,共3页Chinese Pharmaceutical Journal

基　　金：江苏省自然科学基金资助项目!(BK97174)

摘　　要：目的　建立测定全血中环孢素A(CsA)浓度的亲环素A结合法。方法　利用重组人亲环素A(rhCyPA)作为结合蛋白 ,3H CsA作示踪剂 ,活性炭吸附分离结合的与游离的两相 ,测定结合相的放射性。结果　方法的有效测定范围为 5 0～ 16 0 0ng·ml- 1 ,批内 (n =8)RSD为 8.2 % ,批间 (n =6 )RSD为 10 .6 % ,标本提取与测定方法的全程回收率为 10 4% ( 2 40ng·ml- 1 )和 10 6 %( 76 4ng·ml- 1 ) ,测定结果与FPIA法高度相关 (r =0 .96 ;P <0 .0 0 1)。对几种CsA类似物与CyPA的亲和力进行测定的结果表明 ,其亲和力CsA >CsC >CsD >CsB。结论　测定CsA浓度的亲环素A结合法灵敏 ,重复性和稳定性好 。OBJECTIVE To develop a specific cyclophilin A binding assay for the determination of cyclosporine A (CsA) in whole blood.METHODS Recombinant hman cyclophilin A (rhCyPA) was used as the binding protein with the use of 3HCsA as the tracer.Charcoal adsorption was used for separating the bound and free tracer.RESULTS The assay could detect CsA over the range of 50～1600 ng·ml -1 .CVs were 10.6% for interasaay and 8.2% for extraassay.The recovery after extraction was 104% for 240 ng·ml -1 and 106% for 764 ng·ml -1 ,respectively.Thirty blood samples taken from renal allograft recipients were measured with cyclophilin A binding assay and FPIA simultaneoasly,the results were related closely with each other (r=0.96;P<0.001).The binding activity of various CsA analogs was shown as following:CsA>CsC>CsD>CsB.CONCLUSION The cyclophilin A binding assay is sensitive,reproducible and stable.It is useful for measuring CsA in whole blood.

关 键 词：环孢素 亲环素A 血药浓度 监测 

分 类 号：R979.5[医药卫生—药品] R969.1[医药卫生—药学]