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1.
Journal of Experimental Hematology ; (6): 1144-1146, 2015.
Artículo en Chino | WPRIM | ID: wpr-274078

RESUMEN

<p><b>OBJECTIVE</b>The purpose of this research was to identify a novel HLA-B allele in Chinese population.</p><p><b>METHODS</b>The HLA typing of bone marrow donors was performed by PCR-SBT. The ambiguous novel HLA allele was confirmed with GSSP method.</p><p><b>RESULTS</b>The sequence of a sample was different from all alleles in the HLA-B databases. The sequence analysis showed that it differed from the closet matching allele HLA-B*13:01:01 at one nucleotide substitution, 137 T > C in Exon2, which resulted in an amino acid change from Phenylalanine (Phe) to Serine (Ser) at codon 22.</p><p><b>CONCLUSION</b>A novel allele was identified and named as HLA-B*13:68 by the WHO Nomenclature Committee.</p>


Asunto(s)
Humanos , Alelos , Pueblo Asiatico , Secuencia de Bases , Antígenos HLA-B , Prueba de Histocompatibilidad , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN
2.
Chinese Journal of Biotechnology ; (12): 106-111, 2007.
Artículo en Chino | WPRIM | ID: wpr-325410

RESUMEN

This study reports the preparation and identification of soluble programmed death-1 (PD-1) ligand-1 (sPD-L1) and its antibodies of mouse origin. Immobilized metal ion affinity chromatography was used to perform on-column refolding with simultaneous purification of denatured sPD-L1, and soluble sPD-L1 with purity of 95% was obtained. The purified sPD-L1 was verified by immunoblotting using a commercial goat-anti-human PD-L1 antibody. An ELISA-based assay showed that it also had high binding activity for its cognate receptor PD-1. Furthermore, mouse anti-sPD-L1 antiserum of high titer was raised using the purified sPD-L1 as an immunogen, and the specific IgG antibodies were purified using sPD-L1-HiTrap affinity chromatography. In addition, a sensitive sandwich ELISA was established using the purified IgG antibodies together with the commercial goat antibodies. In conclusion, the preparation of soluble sPD-Ll and its antibodies provide the basis for detection of the potential anti-PD-L1 antibodies and soluble PD-L1 in humans as well as for further investigation of its in vivo bioactivities and characterization of its potential receptors.


Asunto(s)
Animales , Femenino , Humanos , Ratones , Anticuerpos , Alergia e Inmunología , Antígenos CD , Genética , Alergia e Inmunología , Metabolismo , Antígeno B7-H1 , Cromatografía de Afinidad , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Escherichia coli , Genética , Sueros Inmunes , Alergia e Inmunología , Immunoblotting , Ratones Endogámicos C57BL , Proteínas Recombinantes , Alergia e Inmunología , Metabolismo , Solubilidad
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