TY - JOUR
T1 - DNA display for in vitro selection of diverse peptide libraries
AU - Yonezawa, Masato
AU - Doi, Nobuhide
AU - Kawahashi, Yuko
AU - Higashinakagawa, Toru
AU - Yanagawa, Hiroshi
N1 - Funding Information:
We thank H. Takashima for critical comments on the manuscript and N. Matsumura for experimental advice. This work was supported in part by a Special Coordination Fund from the Ministry of Education, Science and Technology, Japan.
PY - 2003/10/1
Y1 - 2003/10/1
N2 - We describe the use of a DNA display system for in vitro selection of peptide ligands from a large library of peptides displayed on their encoding DNAs. The method permits completely in vitro construction of a DNA-tagged peptide library by using a wheat germ in vitro transcription/translation system compartmentalized in water-in-oil emulsions. Starting with a library of 10(9)-10(10) random decapeptides, 21 different peptide ligands were isolated for monoclonal antibody anti-FLAG M2. DNA display selected more diverse peptides with a DYKXXD consensus motif than previously reported phage display systems. Binding and recovery rates of three peptides were significantly higher than those of the original FLAG peptide, implying that these peptides would be superior to the FLAG peptide for purification of tagged proteins. The simplicity of DNA display enables two selection rounds per day to be conducted. Further, DNA display can overcome the limitations of previous display technologies by avoiding the use of bacterial cells and RNA tags. Thus, DNA display is expected to be useful for rapid screening of a wide variety of peptide ligands for corresponding receptors.
AB - We describe the use of a DNA display system for in vitro selection of peptide ligands from a large library of peptides displayed on their encoding DNAs. The method permits completely in vitro construction of a DNA-tagged peptide library by using a wheat germ in vitro transcription/translation system compartmentalized in water-in-oil emulsions. Starting with a library of 10(9)-10(10) random decapeptides, 21 different peptide ligands were isolated for monoclonal antibody anti-FLAG M2. DNA display selected more diverse peptides with a DYKXXD consensus motif than previously reported phage display systems. Binding and recovery rates of three peptides were significantly higher than those of the original FLAG peptide, implying that these peptides would be superior to the FLAG peptide for purification of tagged proteins. The simplicity of DNA display enables two selection rounds per day to be conducted. Further, DNA display can overcome the limitations of previous display technologies by avoiding the use of bacterial cells and RNA tags. Thus, DNA display is expected to be useful for rapid screening of a wide variety of peptide ligands for corresponding receptors.
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U2 - 10.1093/nar/gng119
DO - 10.1093/nar/gng119
M3 - Article
C2 - 14500846
AN - SCOPUS:0141905882
SN - 0305-1048
VL - 31
SP - e118
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 19
ER -