A novel one-step strategy for the preparation of HLA/HBc(18-27) and HLA/CEA (694-702) complexes with ion exchange chromatography.
The heavy chain protein of HLA-peptide complexes (HLA/HBc(18-27) and HLA/CEA(694-702)) immobilized onto an ion exchange chromatography column and then the dilution-refolded HBc(18-27)-fused or CEA(694-702)-fused β2m protein was able to pass through the column. Using this method, HLA/peptide complexes were prepared within 30 h with a refolding yield of at least 20% (w/w) and purity of over 80% (w/w). This strategy refolds, concentrates, and purifies HLA/peptide complexes in a single integrated step and offers a potential tool to refold multiple-subunit proteins other than the major histocompatibility complex (MHC)/peptide complexes.
Meng
Fanyan
F
Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, School of Medicine, Southeast University, 210009, Nanjing, Jiangsu, China.
Shen
Chuanlai
C
Miao
Fengqin
F
He
Yong
Y
Shen
Yuqing
Y
Xie
Wei
W
Zhang
Jianqiong
J
D002852:Chromatography, Ion Exchange / Q000379:methods; D015234:HLA-A Antigens / Q000302:isolation & purification* / Q000378:metabolism; D015789:HLA-A2 Antigen; D055503:Protein Multimerization; D058849:Protein Refolding; D001613:beta 2-Microglobulin / Q000302:isolation & purification* / Q000378:metabolism
D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D001613:beta 2-Microglobulin