呋喃唑酮代谢物人工抗原的合成及抗体的制备

Synthesis of artificial antigens and preparation of specific antisera against 3-amino-2-oxazolidinone，a metabolite of furazolidone

关键词:  呋喃唑酮代谢物  特异性抗体  ELISA

Key words:  Metabolite of Furazolidone  Specific antisera  IC-ELISA

摘要:

为进一步研究和完善免疫检测呋喃唑酮代谢物方法，制备了抗呋喃唑酮代谢物（AOZ）特异性抗体。采用对醛基苯甲酸对AOZ进行衍生化得到CPAOZ，还原CPAOZ结构中的C=N双键得到半抗原，半抗原用N-羟基琥珀酰亚胺活化酯法与BSA偶联制备免疫原，免疫新西兰大白兔制得特异性抗体，采用间接（竞争）ELISA法评价抗血清效价及特异性。结果显示，试验获得了较高效价的抗AOZ血清，其效价为1∶1280000，AOZ半抑制浓度为5.9 ng/mL；抗血清与结构类似物呋喃它酮代谢物的交叉反应率仅为0.76%，与呋喃妥因代谢物和呋喃西林代谢物无交叉反应；利用该抗体建立的间接竞争ELISA检测法，AOZ在1～27 ng/mL与抑制率呈线性关系。结果表明，获得的特异性抗体虽然灵敏度较低，却对AOZ而非AOZ衍生物有特异性，可为研究畜产品中AOZ残留检测提供了新的思路和方法。

Abstract:

In order to prepare specific antibody against 3-amino-2-oxazolidinone (AOZ), a metabolite of furazolidone, AOZ was activated by 4-formylbenzoic acid and then reduced the schiff base, then conjugated with BSA via EDC/NHS as immunogen. Rabbits were immunized with immunogen; antisera were evaluated by indirect (competitive) ELISA (IC-ELISA). Antisera titer was 1∶1280000 and IC50 was 5.9 ng/mL. The antisera had hardly cross-reaction with AMOZ, AHD and SEM. An IC-ELISA was developed for the determination of AOZ in the linear range of 1~27 ng/mL. Although the sensitivity is not high, the new antibodies with high specificity against AOZ not AOZ derivatives were obtained in this study which would provide a new thought and method for the detection of AOZ in livestock and poultry products.