Cho Young-Jin, Lee Dae-Hee, Kim Dae-Ok, Min Won-Ki, Bong Ki-Tae, Lee Gang-Gweon, Seo Jin-Ho
School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul 151-742, Korea.
J Agric Food Chem. 2005 Nov 2;53(22):8447-51. doi: 10.1021/jf051681q.
A monoclonal antibody (Mab) against ochratoxin A (OTA) was produced from the hybridoma cell line C7G25, which was established by the fusion of Sp2/0-Ag14 myeloma cells with spleen cells isolated from a BALB/c mouse immunized with the OTA-bovine serum albumin conjugate. This Mab belongs to the IgG(2a) heavy-chain subclass with a kappa-type light chain. The level of 50% inhibition concentration was 1.20 ng/mL in a competitive direct enzyme-linked immunosorbent assay (cdELISA), and the detection limit was 0.12 ng/mL. This antibody is specific for OTA but also shows cross-reactivity with ochratoxin B (31.7%) in a cdELISA. On the basis of the sandwich format using the produced Mab against OTA, a rapid immunochromatographic assay was developed to efficiently detect OTA. This method was able to detect up to 500 ng/mL of OTA in <10 min.
一种抗赭曲霉毒素A(OTA)的单克隆抗体(Mab)由杂交瘤细胞系C7G25产生,该细胞系通过将Sp2/0-Ag14骨髓瘤细胞与从用OTA-牛血清白蛋白偶联物免疫的BALB/c小鼠分离的脾细胞融合而建立。这种单克隆抗体属于具有κ型轻链的IgG(2a)重链亚类。在竞争性直接酶联免疫吸附测定(cdELISA)中,50%抑制浓度水平为1.20 ng/mL,检测限为0.12 ng/mL。该抗体对OTA具有特异性,但在cdELISA中也显示出与赭曲霉毒素B的交叉反应性(31.7%)。基于使用所产生的抗OTA单克隆抗体的夹心形式,开发了一种快速免疫色谱测定法以有效检测OTA。该方法能够在不到10分钟的时间内检测高达500 ng/mL的OTA。