New perspective for phage display as an efficient and versatile technology of functional proteomics |
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Authors: | Wei Li Nora B Caberoy |
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Institution: | (1) Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, 1638 N.W. 10th Avenue, Miami, FL 33136, USA |
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Abstract: | Phage display with antibody libraries has been widely used with versatile applications. However, phage display with cDNA libraries
is rare and inefficient. Because of uncontrollable reading frames and stop codons in cDNA repertoires, high percentage of
phage clones identified from conventional cDNA libraries are non-open reading frames (non-ORFs) encoding unnatural short peptides
with minimal implications in protein networks. Consequently, phage display has not been used as a technology of functional
proteomics to elucidate protein–protein interactions like yeast two-hybrid system and mass spectrometry-based technologies.
Several strategies, including C-terminal display and ORF cDNA libraries, have been explored to circumvent the technical problem.
The accumulative endeavors eventually led to the efficient elucidation of a large number of tubby- and phosphatidylserine-binding
proteins in recent studies by ORF phage display with minimal reading frame issue. ORF phage display inherits all the versatile
applications of antibody phage display, but enables efficient identification of real endogenous proteins with efficiency,
sensitivity, and accuracy comparable to other technologies of functional proteomics. Its ELISA-like procedure can be conveniently
adapted by individual laboratories or fully automated for high-throughput screening. Thus, ORF phage display is an efficient,
sensitive, versatile, and convenient technology of functional proteomics for elucidation of global and pathway-specific protein–protein
interactions, disease mechanisms, or therapeutic targets. |
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