Phage display screening in low dielectric media.
Here we report the first application of phage display screening in low dielectric media. Two series of phage clones with affinity for alpha-chymotrypsin (CT) were selected from a Ph.D.(TM)-C7C library, using either a buffer or acetonitrile in buffer (50%, v/v). The affinity of lysates, individual clones or selected cyclic peptides for the enzyme was studied by examining their influence on CT activity. Peptides displayed on phage selected in buffer provided significant protection from enzyme autolysis resulting in marked increase in CT activity (>100%). Phage selected in ACN provided some, albeit weak, protection from the detrimental influence on CT from ACN. In conclusion, the results demonstrate the potential for the application of phage display screening protocols to targets in media of low dielectricity.
Olofsson
Linus
L
Bioorganic and Biophysical Chemistry Laboratory, School of Pure and Applied Natural Sciences, University of Kalmar, Kalmar, Sweden.
Söderberg
Pernilla
P
Ankarloo
Jonas
J
Nicholls
Ian A
IA
D000465:Algorithms; D000595:Amino Acid Sequence; D017104:Bacteriophage M13 / Q000235:genetics / Q000378:metabolism; D002021:Buffers; D002384:Catalysis / Q000187:drug effects; D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004553:Electric Conductivity; D004789:Enzyme Activation / Q000187:drug effects; D008954:Models, Biological; D019151:Peptide Library; D017186:Titrimetry / Q000379:methods
D002021:Buffers; D019151:Peptide Library; D002918:Chymotrypsin; C117608:alpha-chymotrypsin