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目的制备抗罗丹明B[9-(2-羧基苯基)-3,6-双(二乙氨基)]小鼠单克隆抗体(mAb),初步鉴定其特性,为罗丹明B快速检测方法的开发奠定基础。方法采用活性酯法使罗丹明B偶联于载体牛血清白蛋白(BSA)和卵清白蛋白(OVA),合成免疫抗原罗丹明B-BSA和检测抗原罗丹明B-OVA。将罗丹明B-BSA免疫Balb/c小鼠,通过杂交瘤技术获得杂交瘤细胞株17F2,并通过对添加回收率的测定初步建立了罗丹明B的快速检测方法。结果将该细胞株注射Balb/c小鼠制备腹水mAb,腹水纯化后效价为1∶1.28×105,属于IgG1型。罗丹明B mAb的灵敏度为0.05μg/L,IC50为0.42μg/L。mAb与丁基罗丹明B、异硫氰酸罗丹明B、罗丹明6G、5-羧基四甲基罗丹明、罗丹明110、丽丝胺罗丹明B、罗丹明123的交叉反应率均小于0.3%。辣椒油中罗丹明B的添加回收率在80.6%～104.4%之间,变异系数在16.0%～28.8%之间。结论制备出的罗丹明B mAb为研制罗丹明B的快速检测方法奠定了基础。
Objective To prepare a monoclonal antibody (mAb) against rhodamine B [9- (2-carboxyphenyl) -3,6- bis (diethylamino)] mouse for preliminary characterization of rhodamine B Development laid the foundation. Methods Rhodamine B was coupled to carrier bovine serum albumin (BSA) and ovalbumin (OVA) by an active ester method. The immunogen Rhodamine B-BSA and the detection antigen rhodamine B-OVA were synthesized. Rhodamine B-BSA was used to immunize Balb / c mice, and the hybridoma cell line 17F2 was obtained by the hybridoma technique. The rapid detection method of rhodamine B was established by measuring the recovery of the addition. Results The cell line was injected with Balb / c mice to prepare ascites mAb. The titer of ascites purified was 1: 1.28 × 105, belonging to IgG1 type. Rhodamine B mAb sensitivity of 0.05μg / L, IC50 of 0.42μg / L. The cross-reaction rates of mAb with butyl rhodamine B, rhodamine B isothiocyanate, rhodamine 6G, 5-carboxytetramethylrhodamine, rhodamine 110, lissamine rhodamine B and rhodamine 123 were all less than 0.3 %. The recoveries of Rhodamine B in chilli oil ranged from 80.6% to 104.4% with a coefficient of variation of 16.0% -28.8%. Conclusion Rhodamine B mAb prepared for the development of rapid detection of rhodamine B laid the foundation.