采用放射免疫分析法对长春花组织培养物中的长春碱进行定量测定。

Quantitative Determination of Vinblastine in Tissue Cultures of Catharanthus roseus by Radioimmunoassay.

作者信息

Hirata K, Kobayashi M, Miyamoto K, Hoshi T, Okazaki M, Miura Y

机构信息

Department of Biochemical Engineering, Faculty of Pharmaceutical Sciences, Osaka University, 1-6, Yamadaoka, Suita, 565, Japan.

出版信息

Planta Med. 1989 Jun;55(3):262-4. doi: 10.1055/s-2006-961999.

DOI:10.1055/s-2006-961999

PMID:17262413

Abstract

A radioimmunoassay has been developed for the quantitative determination of small amounts of vinblastine in tissue cultures of CATHARANTHUS ROSEUS (L.) G. Don. Antibody was obtained by the immunization of rabbits against a conjugate of vinblastine with bovine serum albumin. The antibody had a high affinity (Ka = 1.2 x 10 (9)l/mol) and specificity for vinblastine. The usable range of standard curve for assay was 0.5-10 ng/ml. Crude alkaloid extracts of tissue cultures could be assayed and many samples could be processed in one time. It was found that the vinblastine contents of multiple shoot cultures were lower than that of intact plants but much higher than that of callus cultures.

摘要

已开发出一种放射免疫分析法，用于定量测定长春花（CATHARANTHUS ROSEUS (L.) G. Don）组织培养物中的少量长春碱。通过用长春碱与牛血清白蛋白的偶联物免疫兔子获得抗体。该抗体对长春碱具有高亲和力（Ka = 1.2 x 10⁹ l/mol）和特异性。该分析方法的标准曲线可用范围为0.5 - 10 ng/ml。组织培养物的粗生物碱提取物可进行分析，并且一次可处理多个样品。结果发现，丛生芽培养物中的长春碱含量低于完整植株，但远高于愈伤组织培养物。