Identification of the Peptides that Bind to Tyrosine Kinase Receptor EphB2 by Phage Display.
The gene encoding the Ig-like domain of tyrosine protein kinase receptor EphB2 was cloned into the expressing vector pET28a. Under induction with IPTG, the positive strain expressed the fusion protein with a hexahistidine tail on the N-terminal. The protein was purified under denaturing conditions using metal chelate chromatography. The purity was up to 94%. The purified-protein-coated ELISA plate was used as target to screen recombinant phages able to bind onto it, and after three rounds of affinity screening, 19 phages that could bind specifically with EphB2 were isolated from a random phage-displayed seven-peptide library. The peptide sequences of the positive phage clones were analyzed.
Zhang
Xiao-Guang
XG
Department of Biochemistry and Molecular Biology, the Fourth Military Medical University, Xi'an 710032, China. bioyao@fmmu.edu.cn
Yao
Li-Bo
LB
Han
Jiong
J
Liu
Xin-Ping
XP
Han
Jing-Tian
JT
Nie
Xiao-Yan
XY
Su
Cheng-ZHi
CZ