Cloning and expression of human immunoglobulin G4 (hIgG4) hinge region cDNA from blood sample

  • D Kernain
  • P Jayanthi
  • R Noordin
  • MT Ong
Keywords: Human IgG4, hinge region, cDNA, peptide, expression.

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 produce a recombinant peptide antigen that can be used to generate monoclonal antibodies specific to hIgG4. 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 was verified by sequencing and cloned into pQE-2 expression vector to construct pQE-2-IgG4. The latter was transformed Escherichia coli M15 cells, recombinant protein expression induced by IPTG and the expression was confirmed by SDS-PAGE and western blot analysis.

Key words: Human IgG4, hinge region, cDNA, peptide, expression.

Published
2013-10-17
Section
Articles

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eISSN: 1684-5315