动物医学进展2025，Vol.46Issue(8)：19-24,6.
山羊白细胞介素6双抗体夹心ELISA检测方法的建立
Development of Double Antibody Sandwich ELISA for the Detection of Goat Interleukin-6
摘要
Abstract
In order to establish the goat interleukin-6(IL-6)double antibody sandwich ELISA method,the purified goat IL-6 recombinant protein was used as immunogen to immunize rabbits(1 mg/rabbit)and Balb/c mice(100 μg/mouse)to prepare rabbit-derived IL-6 capture polyclonal antibody and mouse-derived IL-6 detection polyclonal antibody,which were used as coating antibody and detection antibody respectively to establish the double antibody sandwich ELISA method.The contents of IL-6 in mammary gland from healthy and mastitis goats were detected using this method.The results showed that the size of the recom-binant IL-6 protein was 38 ku,and the protein had good reactogenicity as identified by Western blot.The coating concentration of coating antibody was 10 μg/mL,and the detection antibody was 1∶800.When the concentration of IL-6 recombinant protein was 0.06 ng/mL,the value of P/N was still greater than 2.1,showing that the sensitivity was good.The detecting result using this method did not differ significantly from the actual concentration,and the inter-and intra-batch coefficients of variations were all below 10％,showing good accuracy and repeatability of this method.Using this method,it was found that IL-6 level was higher in the mammary gland of mastitis goats compared with healthy controls(P＜0.01).Therefore,we established a double antibody sandwich ELISA method for the detection of IL-6,providing a certain techni-cal support for further monitoring goat mastitis.关键词
山羊/白细胞介素6/免疫/双抗体夹心ELISAKey words
Goat/Interleukin-6/Immunization/Double antibody sandwich ELISA分类
农业科技引用本文复制引用
王天星,田曦庆,乔玉娥,张丹辉,何海军,陈德坤,马文涛..山羊白细胞介素6双抗体夹心ELISA检测方法的建立[J].动物医学进展,2025,46(8):19-24,6.基金项目
中央高校基本科研业务费专项资金(2452023072) （2452023072）
