Selecting peptide ligands of microcystin-LR from phage displayed random libraries.
In the present study, we investigated to find novel ligands for low molecular weight environmental toxin, microcystin-LR (MC-LR) by using phage display technology. Two random libraries, displaying linear 12-mer peptides and cyclic 7-mer peptides, were screened against the immobilized target respectively. After three rounds of panning, phage clones that recognized microcystin-LR specifically were obtained from both the linear and the constrained libraries, proved by enzyme-linked immumosorbent assays and immunoprecipitation assays. DNA sequencing indicated that peptides displayed on some of the selected clones shared consensus sequences. Compared with traditional methods, this approach provided a cheaper and more rapid alternative to screen specific ligands for microcystin-LR. Moreover, since it is rather difficult to take small molecules as targets of phage display libraries, the success of this experiment expanded the applications of phage display technology, and provided a new avenue to study environmental small molecular toxins.
Zhao
S W
SW
Department of Biochemistry, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China.
Shen
P P
PP
Zhou
Y
Y
Wei
Y
Y
Xin
X B
XB
Hua
Z C
ZC
D001427:Bacterial Toxins; D004797:Enzyme-Linked Immunosorbent Assay; D047468:Immunoprecipitation; D008024:Ligands; D008387:Marine Toxins; D052998:Microcystins; D019151:Peptide Library; D010456:Peptides, Cyclic / Q000737:chemistry*; D017422:Sequence Analysis, DNA
D001427:Bacterial Toxins; D008024:Ligands; D008387:Marine Toxins; D052998:Microcystins; D019151:Peptide Library; D010456:Peptides, Cyclic; C057862:cyanoginosin LR