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.
出处
《中国药学杂志》
CAS
CSCD
北大核心
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)
关键词
环孢素
亲环素A
血药浓度
监测
cyclosporine A
cyclophilin A
therapeutic drug monitoring
