玉米赤霉烯酮多克隆抗体的制备及特性分析

合成玉米赤霉烯酮半抗原,应用液相色谱-质谱联用法进行鉴定,并采用活泼酯法将半抗原与载体蛋白OVA或BSA偶联,分别作为免疫原或包被原,并通过SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和紫外扫描法鉴定偶联效果,免疫3只BALB/c小鼠,制备玉米赤霉烯酮多克隆抗体。结果表明:抗血清效价最高达1∶32 000,以此多抗建立的玉米赤霉烯酮间接竞争标准曲线IC50为39.8ng/mL,IC10为0.71ng/mL,多抗与玉米赤霉烯酮类似物β-zearalenol、zear-alanone、α-zearalanol、β-zearalanol交叉反应率分别为4.80%,3.07%,0.96%,0.09%。说明试验成功制备了玉米赤霉烯酮人工抗原及高特异性多克隆抗体。

Preparation and characterization of polyclonal antibodies against zearalenone

The hapten of zearalenone with carboxyl group was synthetized,and then identified using liquid chromatography/mass spectrometry.The hapten was coupled with carrier protein ovabumin(OVA) or bovine serum albumin(BSA) to prepare artificial antigen by active ester method.And then,was identified by SDS-PAGE and UV scanning spectrum.Three mice were immunized to prepare polyclonal antibody of zearalenone.Titres of antibodies were determined to be 1∶32 000 by indirect ELISA,and the antibodies showed high sensitivity with IC50 value of 39.8 ng/mL toward zearelanone.Cross-reactivities with analogues were as follows: β-zearalenol,4.8%;zearalanone,3.07%;α-zearalanol,0.96%;β-zearalanol,0.09%;The results show that the artifical antigens and polyclonal antibody of zearalenone were developed successfully.