Cloning of pCDNA3-IgG4 and pQE-2-IgG4 human hinge region cDNA for intrasplenic and intraperitoneal immunization

Abstract

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.

Key words: Component, human IgG4, hinge region, cDNA, vector, expression.