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Title 

Generation of anti-HLA-DR4 specific antibodies by immunization of the recombinantly expressed allelic subtype-specific region of the HLA-DRB1 0405 molecules

Authors 

Jung Hyun ParkEun Wie ChoYun Jung LeeJin ChungKyung Soo HahmKil Lyong Kim

Publisher 

Springer Verlag (Germany)

Issue Date 

1998

Citation 

Journal of Biochemistry and Molecular Biology, vol. 31, no. 2, pp. 111-116

Keywords 

Allelic typingAntibodiesHLA-DRMHC Class IIRecombinant protein

Abstract 

HLA-DR4 is the dominant allele of MHC class II genes in Koreans. In particular, the DRB1*0405 subtype has been reported to be almost exclusively expressed in Far East Asians, and has also been observed to be strongly associated with rheumatoid arthritis in Koreans and the Japanese. Identification of this specific allele has been mainly performed by PCR-based methods, which is often time consuming, costly, and involves tedious procedures such as the isolation of genomic DNA, PCR, and gel electrophoresis. To develop a more convenient tool for screening vast amounts of samples as well as to generate reagents which might also be used in other applications, in this study, antibodies were produced against this specific HLA subtype. By PCR, an allele-specific region covering the β1 domain of DRB1*0405 was amplified and recombinantly expressed in E.coli. Immunization of Lewis rats with the purified protein yielded an allele specific antiserum. Western blot analysis showed the selective detection of the HLA-DR β-chain. Using this antiserum, established cell lines and peripheral blood lymphocytes were analyzed on their HLA haplotype by fluorescence activated flow cytometry. These novel antibodies will provide a powerful tool in the detection and investigation of DR4 alleles.

ISSN 

1225-8687

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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