一种水溶性免疫佐剂应用于小分子化合物单克隆抗体制备的效果评价（英文）

Evaluation of a water-soluble adjuvant for the development of monoclonal antibodies against small-molecule compounds

目的：评价一种新型水溶性免疫佐剂在制备小分子化合物单克隆抗体（单抗）中的应用效果。创新点：采用一种新型水溶性免疫佐剂，快速制备了分别针对4种农药和1种海洋毒素的高亲合力单抗，且其性能与以往采用弗氏佐剂获得的相应单抗非常接近。方法：本研究以4种农药和1种海洋毒素为目标分析物，采用一种新型水溶性佐剂与小分子抗原相互混合，直接对Balb／c小鼠进行小腿肌肉注射免疫。经过2～3次免疫后进行腹腔注射常规末次免疫，细胞融合后筛选出了相应的杂交瘤细胞株，制备并鉴定了针对上述半抗原小分子的特异性单抗。结论：采用该水溶性免疫佐剂制备抗原注射溶液无需乳化过程，免疫操作简便，且对小鼠负面效应较小；虽然该处理小鼠产生的抗血清效价并没有像常规弗氏免疫佐剂带来的效价值高，但细胞融合后筛选得到有效细胞株的概率尚可；获得的针对目标分析物的单抗在灵敏度和特异性等方面都接近于以往采用弗氏佐剂获得的单抗。试验结果表明，该水溶性免疫佐剂适用于针对小分子化合物单抗的高效制备。

A water-soluble adjuvant named QuickAntibody （QA） was introduced into the procedure of mouse immunization for the development of hapten-specific monoclonal antibodies （mAbs）, using four kinds of pesticides as model compounds. Compared with conventional Freund＇s adjuvants, QA treatments offered relatively low but acceptable antiserum titers after three inoculations, gave little adverse effects to the experimental animals, and were preferable in harvesting splenocytes during the steps of cell fusion. Afterwards, hybridomas from the QA group were prepared and screened by both non-competitive and competitive indirect enzyme-linked immunosorbent assays （ELISAs）. The efficiency of gaining immune-positive hybridomas was satisfactory, and the resultant mAbs showed sensitivities （half maximal inhibitory concentration （IC50）） of 0.91, 2.46, 3.72, and 6.22 ng/ml to triazophos, parathion, chlorpyrifos, and fenpropathrin, respectively. Additionally, the performance of QA adjuvant was further confirmed by acquiring a high-affinity mAb against okadaic acid （IC50 of 0.36 ng/ml） after three immunizations. These newly developed mAbs showed similar or even better sensitivities compared with previously reported mAbs specific to the corresponding analytes. This study suggested that the easy-to-use adjuvant could be applicable to the efficient gen- eration of highly sensitive mAbs against small compounds.