中国免疫学杂志2016，Vol.32Issue(9)：1333-1337,5.DOI:10.3969/j.issn.1000-484X.2016.09.019
酵母表面展示多肽片段分析Δ42 PD1的免疫原性
Analysis of immunogenicity ofΔ42PD1 via yeast surface displaying peptide frag-ments
摘要
Abstract
Objective:To analyze the immunogenicity of the extracellular region of Δ42PD1.Methods: Six fragments ofΔ42PD1 extracellular region-encoding sequence were amplified by PCR, and were cloned into pCTCON2 vector, a yeast surface displaying vector.Yeast cells were transfected with Δ42PD1 fragment-carrying plasmids, then yeast cells were spread on SDCAA plates.Single cell clones were selected and cultured in SGCAA media to induce expression of the target genes.Mouse anti-humanΔ42PD1 anti-serum were generated by immunization of BALB/c mice via intramuscular injection ofΔ42PD1-carrying plasmid plus in-situ electroporation.The binding of anti-serum with yeast cells surface-displaying Δ42PD1 fragments were analyzed using flowcytometry.Results:Nucleotide sequences analysis indicated that the amplified six fragments ofΔ42PD1 sequence length were 110 bp,and the isolated sequence ofΔ42PD1 fragments were 100%homology with PD1 gene previously registered in GenBank.Results from flowcytometry showed that among the six fragments of Δ42PD1 displaying on the surface of yeast cells,F3 and F2 profoundly boundΔ42PD1-specific polyclonal antibodies.Conclusion:F3 and F2 ofΔ42PD1 is an immunogenic dominant region,which pave the way for generation of Δ42PD1-specific monoclonal antibody and epitope mapping.关键词
程序性细胞死亡受体1/Delta-42程序性细胞死亡受体1/酵母表面展示/免疫原性/流式细胞术Key words
PD1/Δ42PD1/Yeast surface display/Immunogenicity/Flowcytometry分类
医药卫生引用本文复制引用
程林,王子乔,徐六妹,唐娴,周泱,王辉..酵母表面展示多肽片段分析Δ42 PD1的免疫原性[J].中国免疫学杂志,2016,32(9):1333-1337,5.基金项目
本文为国家自然科学基金（31500750）、广东省自然科学基金（2016A030313030）和深圳市科技创新基金（JCYJ20140411111718162，JCYJ20150402111430650）资助项目。 （）
