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目的合成可溶性HLA-G1-肽复合物。方法利用原核高效表达的可溶性HLA-G1重链和轻链(β2m),在人工合成的HLA-G1限制性九肽(KGPPAALTL)存在的条件下,通过稀释复性折叠成可溶性HLA-G1-肽复合物。利用特异性抗体(W6/32及兔抗人β2m抗体)进行免疫印迹及酶联免疫吸附试验,检测稀释复性的折叠产物。结果折叠复合物含有35%可溶性HLA-G1-肽复合物、5%可溶性HLA-G1重链聚合体及60%复性的β2m3种成分。结论通过原核表达、体外折叠能够获得大量可溶性HLA-G1-肽复合物。
Objective To synthesize soluble HLA-G1-peptide complexes. Methods The soluble HLA-G1 heavy chain and light chain (β2m) highly expressed in prokaryotic cells were folded into soluble HLA-G1-peptides by dilution refolding in the presence of synthetic HLA-G1-restricted nonapeptide (KGPPAALTL) Complex. Western blotting and enzyme-linked immunosorbent assay (ELISA) were performed using specific antibodies (W6 / 32 and rabbit anti-human β2m antibody) to detect the refolding dilution product. Results The folded composite contained 35% soluble HLA-G1-peptide complex, 5% soluble HLA-G1 heavy chain polymer and 60% refolded β2m3 components. Conclusions A large number of soluble HLA-G1-peptide complexes can be obtained by prokaryotic expression and in vitro folding.