A High-Throughput Phage Display Screening Method Using a Combination of Real-Time PCR and Affinity Chromatography

Title: A High-Throughput Phage Display Screening Method Using a Combination of Real-Time PCR and Affinity Chromatography

Volume: 9 Issue: 1

Author(s): Kengo Morohashi, Tsuyoshi Arai, Seiich Saito, Madoka Watanabe, Kengo Sakaguchi and Fumio Sugawara

Affiliation:

Keywords: Phage display, affinity chromatography, real time PCR, drug target, target validation, NK109

Abstract: Phage display is one of the best methods to identify drug targets, although technical problems including imprecision in quantifying phage and false-positive results are common. To address these difficulties, we propose two methods to more rapidly identify drug-binding phage particles. First, quantification of phage using SYBR Green Realtime PCR significantly improved accuracy and reproducibility. Second, affinity-column chromatography for selection of drug-binding phage particles concentrated particles more than a 96-well plate, making a phage amplification step, which can bias phage distribution, unnecessary. The methods proposed here should be suitable for high-throughput phagedisplay screenings and ultimately lead to more rapid identification of drug targets.

Cite this article as:

Morohashi Kengo, Arai Tsuyoshi, Saito Seiich, Watanabe Madoka, Sakaguchi Kengo and Sugawara Fumio, A High-Throughput Phage Display Screening Method Using a Combination of Real-Time PCR and Affinity Chromatography, Combinatorial Chemistry & High Throughput Screening 2006; 9 (1) . https://dx.doi.org/10.2174/138620706775213840