Kido Katsumi, Morinaga Osamu, Shoyama Yukihiro, Tanaka Hiroyuki
Department of Phamacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan.
Talanta. 2008 Oct 19;77(1):346-50. doi: 10.1016/j.talanta.2008.06.034. Epub 2008 Jul 1.
To establish an immunoassay for baicalin (BA), a hybridoma cell line (9D6) secreting a monoclonal antibody (MAb) against BA was prepared by cell fusion with splenocytes derived from a mouse immunized with BA-bovine serum albumin (BSA) conjugate and a myeloma cell line, SP2/0-Ag14. MAb 9D6 shows specific reactivity against BA and its aglycone, baicalein, but not against other natural products. We developed an enzyme-linked immunosorbent assay (ELISA) using MAb 9D6 in a competitive manner, ranging from 200 ng/mL to 2 microg/mL. After validating the developed ELISA on the basis of intra- and inter-assays and a recovery experiment, it was found that the ELISA was not only simple, but also sufficiently reliable and accurate for quality control of Scutellariae Radix. It allowed determination of BA in complex and mixed materials, such as Kampo medicines.
为建立一种黄芩苷（BA）免疫测定法，通过将用BA-牛血清白蛋白（BSA）偶联物免疫的小鼠脾细胞与骨髓瘤细胞系SP2/0-Ag14进行细胞融合，制备了分泌抗BA单克隆抗体（MAb）的杂交瘤细胞系（9D6）。单克隆抗体9D6对BA及其苷元黄芩素表现出特异性反应，但对其他天然产物无反应。我们以竞争方式使用单克隆抗体9D6开发了一种酶联免疫吸附测定法（ELISA），检测范围为200 ng/mL至2 μg/mL。在基于批内和批间测定以及回收率实验对所开发的ELISA进行验证后，发现该ELISA不仅简单，而且对于黄芩质量控制而言足够可靠和准确。它能够测定复杂和混合材料中的BA，如汉方药物。
