| 磺胺嘧啶单克隆抗体的制备与鉴定 |
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| 引用本文： | 王刘花,许 杨,郭杰标,张 泓,何庆华. 磺胺嘧啶单克隆抗体的制备与鉴定[J]. 食品科学, 2010, 31(11): 192-196. DOI: 10.7506/spkx1002-6630-201011041 |
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| 作者姓名： | 王刘花 许 杨 郭杰标 张 泓 何庆华 |
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| 作者单位： | 南昌大学 食品科学与技术国家重点实验室，中德联合研究院 |
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| 基金项目： | 江西省科技支撑计划项目(2007BS12501) |
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| 摘 要： | 采用重氮法同时制备结合比分别为9.0、8.0、3.7 的3 种磺胺嘧啶免疫抗原，并将其免疫BALB/c 小鼠，其中结合比为8.0 的一组免疫效果最佳。获得一株稳定分泌抗磺胺嘧啶单克隆抗体的杂交瘤细胞(1F6)，体内诱生法制备腹水。间接ELISA 测定腹水效价为1:1.28 × 106，建立间接竞争ELISA 标准曲线，其检测下限为10.4ng/mL，线性范围为14.8～421.5ng/mL，IC50 值为79.1ng/mL，与磺胺二甲嘧啶、磺胺间甲氧嘧啶、磺胺对甲氧嘧啶交叉反应率分别为 0.3%、0.99%、4.5%，与磺胺甲噁唑、磺胺喹恶啉、对氨基苯甲酸、磺胺、对氨基苯磺酸未见有交叉反应。经鉴定，单克隆抗体免疫球蛋白为鼠IgG 的2b 亚类。
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| 关 键 词： | 磺胺嘧啶 重氮法 酶联免疫检测 单克隆抗体 |
| 收稿时间： | 2009-11-09 |
Preparation and Identification of Anti-sulfadiazine Monoclonal Antibody |
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| WANG Liu-hua,XU Yang,GUO Jie-biao,ZHANG Hong,HE Qing-hua. Preparation and Identification of Anti-sulfadiazine Monoclonal Antibody[J]. Food Science, 2010, 31(11): 192-196. DOI: 10.7506/spkx1002-6630-201011041 |
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| Authors: | WANG Liu-hua XU Yang GUO Jie-biao ZHANG Hong HE Qing-hua |
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| Affiliation: | Sino-Germany Joint Research Institute, State Key Laboratory of Food Science and Technology, NanchangUniversity, Nanchang 330029, China |
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| Abstract: | In order to generate a monoclonal antibody (McAb) against sulfadianzine (SD), three separate immunogens with SD/BSA conjugation ratios of 9.0, 8.0 and 3.7 were prepared by diazotization method and BALB/c mice were then immunized with the immunogens. An optimum immunization was observed in BALB/c mice immunized with the immunogen with middle SD/BSA conjugation ratio. A McAb (1F6) with high affinity to SD was obtained by hybridoma technique. The titer of the McAb was 1:1.28 ×106 in ascites according to the indirect ELISA determination. IC50 and the lowest detectable limit (LDL) of the McAb in linear range of 14.8－421.5 ng/mL for SD were 79.1 ng/mL and 10.4 ng/mL, respectively. The cross-reactivity ratios of the McAb against sulfamethazine, sulfamonomethoxine and sulfamethoxydiazine were 0.3%, 0.99% and 4.5%, respectively, while no cross-reactivity to sulfamethoxazole, sulfaquinoxaline, 4-aminobenzoic acid, sulfanilamide or sulfanilic acid was observed. The sub-class of the McAb was identified as IgG2b. |
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| Keywords: | sulfadiazine diazotization ELISA monoclonal antibody |
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