摘要： 以戊二醛法将氨苄青霉素(Ampicillin,Amp)偶联于牛血清白蛋白(BSA)和卵清白蛋白(OVA),合成免疫抗原BSA-Amp和检测抗原0VA-Amp,并进行了鉴定;用BSA-Amp免疫BALB/c小鼠,应用杂交瘤技术建立了株分泌抗Amp单克隆抗体的杂交瘤细胞株.ELISA和竞争ELISA结果显示,单克隆抗体1G7、2A11、2D10和3F6的腹水抗体效价分别为1:2.56×107、1:1.28×107、1:5.12 ×106和1:5.12×106,Amp的半数抑制浓度(IC50)分别为10.22,3.58,.03,.95 ng/mL.Amp单克隆抗体与羧苄青霉素(1G7、2A11、2D10和3F6)、青霉素G(1G7和3F6)、阿莫西林(1G7)等青霉素类抗生素存在显著的交叉反应,而与双氯霉素和邻氯霉素未见明显交叉反应.
关键词:
氨苄青霉素,
单克隆抗体,
交叉反应
Abstract: Ampicillin(Amp) was coupled to carrier proteins of bovine serum albumin(BSA) and ovalbumin(OVA) to synthetize immunogen BSA-Amp and detection antigen OVA-Amp.Four hybridoma cell lines secreting monoclonal antibodies(mAbs) specific for Amp were established by immunizing BALB/c mice with BSA-Amp using hybridoma technology.The antibody titers of mAb ascites(1G7,2A11,2D10 and 3F6) were determined as 1∶2.56×107,1∶1.28×107,1∶5.12×106 and 1∶5.12×106 in ELISA,and their half maximal inhibitory concentration(IC50) of Amp were 10.22,3.58,4.03,4.95 ng/mL in the inhibitive ELISA,respectively.The Amp mAbs showed significant cross-reactivity to the penicillin antibiotics of carbenicillin(1G7,2A11,2D10 and 3F6),penicillin G(1G7 and 3F6),and amoxicillin(1G7),whereas no obvious cross-reactivity to dicloxacillin or cloxacillin was found.
Key words:
Ampicillin,
Monoclonal antibody,
Cross-reactivity
中图分类号:
李青梅, 刘庆堂, 王寅彪, 柴书军, 王自良, 郭军庆, 职爱民, 孟红丽, 张改平. 氨苄青霉素单克隆抗体及其与青霉素类抗生素交叉反应性[J]. 华北农学报, 2011, 26(3): 60-63. doi: 10.7668/hbnxb.2011.03.013.
LI Qing-mei, LIU Qing-tang, WANG Yin-biao, CHAI Shu-jun, WANG Zi-liang, GUO Jun-qing, ZHI Ai-min, MENG Hong-li, ZHANG Gai-ping. Anti-ampicillin Monoclonal Antibodies and Their Cross-reactivity to Penicillin Antibiotics[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(3): 60-63. doi: 10.7668/hbnxb.2011.03.013.