Application of Beta2-microglobulin-ZsGreen in binding affinity assay of MHC- Ⅰ peptide
XIONG Ruihua
WANG Haoliang
ZOU Liyun
LI Jingyi
ZHANG Jingyu
ZHANG Ronghua
WAN Ying
Abstract：Objective To establish a novel and convenient MHC-Ⅰ peptides binding affinity assay by using recombinant β2-microglobulin-ZsGreen fusion protein. Methods An eukaryotic expression vector was constructed by linking the sequence of theβ2-mi-croglobulin and ZsGreen-6×His Tag employing molecular cloning technique. Theβ2-microglobulin-ZsGreen fusion protein was ex- pressed in 293FTclis, and then purified with Ni-affinity chromatography. The fusion protein was co-cuhured with T2 cell and positive MHC- Ⅰ binding epitopes to establish the novel MHC- Ⅰ peptides binding affinity assay. Results β2-microglobulin-ZsGreen fusion protein was obtained and the convenient MHC- Ⅰ peptides binding affinity assay was established. Conclusion The newly established MHC- Ⅰ peptides binding affinity assay for CTL epitope identification is a feasible and simple method, which can be used in large flux epitope identification.
Keywords：green fluorescent proteinsfusion proteinsaffinity labelsT-cell epitopes
Publication Date：2009-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 452-456 )