利用针对凝集素A亚基的单克隆抗体研究槲寄生制剂中凝集素的异质性。

Study of heterogeneity of lectins in mistletoe preparations by monoclonal antibodies to their A-subunits.

作者信息

Tonevitsky A G, Agapov I, Temiakov D, Moisenovich M, Maluchenko N, Solopova O, Würzner G, Pfueller U

机构信息

Institute of Transplantology and Artificial Organs, Moscow, Russia.

出版信息

Arzneimittelforschung. 1999 Nov;49(11):970-5. doi: 10.1055/s-0031-1300535.

DOI:10.1055/s-0031-1300535

PMID:10604051

Abstract

Monoclonal antibodies (monAbs) displaying diverse affinity to native and recombinant mistletoe lectins A-chains (ML I-A, ML II-A, ML III-A and rML-A) have been obtained. In accordance to specificity of monAb they have been classified into three groups: 1. monAb against MLI-A and MLII-A, 2. monAb against MLII-A and MLIII-A, 3. monAb against A-subunits of MLI, MLII and MLIII. The results indicate, that antigen determinants of mistletoe lectins recognized by monAb MNA4, MNA9 and MTC12 do not contain any carbohydrates. Assay of lectins in mistletoe preparations was based on enzyme-linked lectin assay to meet the requirements given in the guidelines for drug tests. In this paper a sandwich ELISA test-system is described which allows to identify each of three ML-toxins. With the detection limits below 3 ng/ml and a linear measuring range of 3-30 ng/ml, dosages of mistletoe lectins in therapeutic range can be assayed. The problems in mistletoe lectins determination, structural differences and nature of heterogeneity of this proteins are discussed.

摘要

已获得对天然和重组槲寄生凝集素A链（ML I-A、ML II-A、ML III-A和rML-A）具有不同亲和力的单克隆抗体（单克隆抗体）。根据单克隆抗体的特异性，它们被分为三组：1. 针对MLI-A和MLII-A的单克隆抗体，2. 针对MLII-A和MLIII-A的单克隆抗体，3. 针对MLI、MLII和MLIII的A亚基的单克隆抗体。结果表明，单克隆抗体MNA4、MNA9和MTC12识别的槲寄生凝集素的抗原决定簇不包含任何碳水化合物。槲寄生制剂中凝集素的测定基于酶联凝集素测定，以满足药物测试指南中的要求。本文描述了一种夹心ELISA检测系统，该系统可以识别三种ML毒素中的每一种。检测限低于3 ng/ml，线性测量范围为3-30 ng/ml，可以测定治疗范围内的槲寄生凝集素剂量。讨论了槲寄生凝集素测定中的问题、这种蛋白质的结构差异和异质性的性质。