伏马菌素B1免疫学检测方法的建立

宫慧芝; 计融; 杨军; 江涛; 李凤琴

doi:10.11847/zgggws2006-22-07-44

伏马菌素B1免疫学检测方法的建立

Establishment of immunoassay for detecting fumonisin B1

摘要

摘要:
目的建立应用单克隆抗体的伏马菌素B₁(fumonisin B₁, FB₁)间接竞争酶联免疫吸附检测方法。
方法利用B淋巴细胞杂交瘤技术制备抗FB₁单克隆抗体, 建立FB₁间接竞争酶联免疫吸附方法。
结果建立稳定分泌抗FB₁毒素单克隆抗体的杂交瘤细胞株, 分泌的抗体为IgG1亚类, 分子量150KD, 腹水稀释度为1:2.0×10⁸, 亲和常数为6.72×10⁹L/mol。FB₁间接竞争酶联免疫吸附方法的最低检出浓度为5μg/L, 校正曲线的线性范围50~500μg/L, 线性方程Y=-0.582X+1.793(r=0.99, P < 0.05);与脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮、T-2毒素均无交叉反应。回收率71.89%~112.95%, 平均回收率100.23%。
结论所建立的检测方法具有快速、简便、灵敏的特点, 可满足实际工作需要。

Abstract:
Objective To develop enzyme-linked immunosorbant assay for detection of FB₁ which is based on monclonal antibodies.
Methods To produce hybridoma cell lines excreting monoclonal antibodies against FB₁ by using B cell hybridomatechnique and develop enzyme-linked immunosorbant assay for detection of FB₁.
Results Hy bridoma cell lines excreting monoclonal antibodies were developed and monoclonal antibodies against FB₁ were prepared.Monoclonal antibodies produced by the hybridoma cells were tested for subtypes and designated as IgG1, the mo lecular weight was 150kD, the titer was 1:2.0~10⁸, the affinity constant was 6.72×10⁹ L/mol.The limited concentration of detection was 5μg/L, linear range was 50~500 μg/L, the linearequation was Y=-0.582x+1.793, (r=0.99, P < 0.05).There was no cross reaction with DON, ZEN, T-2 toxin.The recovery rate of spiked maize was among 71.89% ~112.95%, and mean recovery rate was 100.23%.
Conclusion The detection method is simple, rapid, sensitive and can meet the demand of practical work.

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