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