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首页> 外文期刊>African Journal of Biotechnology >Cloning of pCDNA3-IgG4 and pQE-2-IgG4 human hinge region cDNA for intrasplenic and intraperitoneal immunization
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Cloning of pCDNA3-IgG4 and pQE-2-IgG4 human hinge region cDNA for intrasplenic and intraperitoneal immunization

机译：pCDNA3-IgG4和pQE-2-IgG4人铰链区cDNA的克隆用于脾内和腹膜内免疫

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Human immunoglobulin G4 (hIgG4) is increasingly being used for?the?detection of various infectious diseases and in allergy-related immunoassays, thus, anti-hIgG4 antibody is of interest in the development of diagnostic tests. The present study was conducted to prepare the plasmid construct of pQE-2-IgG4 for peptide expression and pCDNA3-IgG4 for use in intrasplenic immunization in view of monoclonal antibody production. pQE-2 is a prokaryotic expression vector whereas pCDNA3 is a mammalian expression vector. Some methods were used to compare the efficacy of the immunization routes in raising monoclonal antibody against human IgG4 in Balb/c mice. The cDNA coding for the hinge region of hIgG4 was derived from mRNA from a human blood sample using reverse transcription polymerase chain reaction (RT-PCR). The cDNA sequence was verified by sequencing. The cDNA fragment was then cloned into pQE-2 and pCDNA3, producing pQE-2-IgG4 and pCDNA3-IgG4, respectively. pQE-2 was transformed into?M15?cell to produce the peptide of interest.
机译：人免疫球蛋白G4（hIgG4）越来越多地用于检测各种传染病以及与变态反应相关的免疫测定，因此，抗hIgG4抗体在诊断测试的开发中引起了人们的兴趣。鉴于单克隆抗体的产生，进行了本研究以制备用于肽表达的pQE-2-IgG4和用于脾内免疫的pCDNA3-IgG4的质粒构建体。 pQE-2是原核表达载体，而pCDNA3是哺乳动物表达载体。一些方法用于比较免疫途径在Balb / c小鼠中产生抗人IgG4单克隆抗体的功效。使用逆转录聚合酶链反应（RT-PCR），从人类血液样本中的mRNA提取编码hIgG4铰链区的cDNA。通过测序验证cDNA序列。然后将cDNA片段克隆到pQE-2和pCDNA3中，分别产生pQE-2-IgG4和pCDNA3-IgG4。 pQE-2被转化入“ M15”细胞以产生目的肽。

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