[1]王莹,郑浩,何成华,等.伏马菌素B1人工抗原的鉴定及抗体的制备[J].南京农业大学学报,2011,34(5):93-98.[doi:10.7685/j.issn.1000-2030.2011.05.017]
 WANG Ying,ZHENG Hao,HE Cheng-hua,et al.Characterization of fumonisin B1artificial antigen and production of the antibody[J].Journal of Nanjing Agricultural University,2011,34(5):93-98.[doi:10.7685/j.issn.1000-2030.2011.05.017]
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伏马菌素B1人工抗原的鉴定及抗体的制备()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
34卷
期数:
2011年5期
页码:
93-98
栏目:
出版日期:
2011-10-18

文章信息/Info

Title:
Characterization of fumonisin B1artificial antigen and production of the antibody
作者:
王莹郑浩何成华张爱华张海彬*
南京农业大学动物医学院,江苏 南京 210095
Author(s):
WANG YingZHENG HaoHE Cheng-huaZHANG Ai-huaZHANG Hai-bin*
College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China
关键词:
伏马菌素B1人工抗原偶联比抗体
Keywords:
fumonisin B1artificial antigencoupling ratioantibody
分类号:
S852.4+3
DOI:
10.7685/j.issn.1000-2030.2011.05.017
文献标志码:
A
摘要:
采用戊二醛(GA)一步法将半抗原伏马菌素B1(FB1)分别与鸡卵清蛋白(OVA)、牛血清白蛋白(BSA)偶联,制备2种具有免疫原性的FB1人工抗原FB1-OVA和FB1-BSA。分别采用凝胶电泳法、紫外光谱法(UV)、红外光谱法(IR)、基质辅助激光分析电离飞行时间质谱法(MALDI-TOF-MS)4种方法鉴定人工抗原偶联效果并测定偶联比。将FB1-BSA作为免疫抗原免疫BALB/c小鼠,FB1-OVA作为固相包被抗原,采用间接酶联免疫吸附法(i-ELISA)测定小鼠抗血清滴度。结果表明:4种方法均鉴定FB1人工抗原合成成功。凝胶电泳法测定FB1-OVA和FB1-BSA相对分子质量分别约为50 000和75 000。MALDI-TOF-MS测定FB1-OVA和FB1-BSA相对分子质量分别为48 009.212和74 355.301,并精确计算FB1-OVA和FB1-BSA偶联比分别为5∶1和10∶1。FB1-BSA免疫后小鼠的抗血清效价为1∶1.25×105,并具有较高灵敏度。
Abstract:
In the study,the hapten fumonisin B1(FB1)was coupled with carrier proteins ovalbumin(OVA)and bovine serum albumin(BSA)respectively,by one step method with glutaraldehyde(GA)method,and two artificial antigens FB1-OVA and FB1-BSA were synthesized.Four methods of gel electrophoresis,ultraviolet spectroscopy(UV),infrared spectroscopy(IR)and matrix-assisted laser absorption ionization time-of-flight mass spectrometry(MALDI-TOF-MS)were used to determine the coupled effects and coupling ratios of the artificial antigens.Then FB1-BSA was used to immunize BALB/c mice,and FB1-OVA was used as solid phase coating antigen.The indirect enzyme-linked immunosorbent assay(i-ELISA)was used to determine the titer of anti-serum from mice.The results showed that four methods all proved that carrier proteins coupled with FB1 successfully.The molecular weights of FB1-OVA and FB1-BSA were about 50 000 and 75 000 respectively by gel electrophoresis.The accurate molecular weights of FB1-OVA and FB1-BSA were 48 009.212 and 74 355.301 and the coupling ratios of FB1-OVA and FB1-BSA were 5∶1 and 10∶1 respectively by MALDI-TOF-MS.The titer of anti-serum was 1∶1.25×105 by i-ELISA with good sensitivity.

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备注/Memo

备注/Memo:
收稿日期:2010-12-22 基金项目:国家863计划项目(2007AA10Z429);江苏省农业三项工程项目(SX(2010)268);江苏省动物源食品与安全保障重点实验室项目(R201017);国家自然科学基金项目(30901091) 作者简介:王莹,博士研究生。*通讯作者:张海彬,教授,从事农畜产品中真菌毒素的检测与去毒研究, E-mail:haibinzh@njau.edu.cn。
更新日期/Last Update: 2011-10-31