期刊
PROTEIN EXPRESSION AND PURIFICATION
卷 166, 期 -, 页码 -出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2019.105520
关键词
Affibody; PD-1/PD-L1; Yeast two hybrid
类别
资金
- Scientific and Technological Research Program of Tianjin Municipal Education Commission
- Foundation of Tianjin Engineering Research Center of Safety Control Technology in Food Processing, P. R. China [GCZX201802]
- Foundation of Tianjin Engineering Research Center of Microbial Metabolism and Fermentation Process Control, P. R. China [ZXKF20180201]
An affibody is a 58 amino acids peptide derived from the Z domain of staphylococcal protein A and generally applied in areas such as imaging diagnosis, clinical therapeutics and biotechnology research. To screen for an affibody targeting the immune checkpoint PD-L1, a combinatorial affibody library was generated in yeast using degenerate overlap PCR primers and In-fusion technology. Z-jl and Z-j2 affibodies targeting the Ig-like V domain of PD-Ll were screened and identified from this combinatorial library using the yeast two hybrid system. The Zji and Z-j2 recombinant affibody proteins were over produced in E.coli and purified. ELISA and GST pull-down assays showed that recombinant Z-jl and Z-j2 affibody proteins bound with high affinity to PD-Ll and inhibited the interaction of PD-1/PD-Ll. Thus, novel affibodies targeting the immune checkpoint PD-1/PD-L1 were identified and produced in this study and have the potential to be used in cancer immunotherapy.
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