DeRisi Lab Phage Immunoprecipitation Sequencing (PhIP-Seq) v1
Year: 2023
Type: preprint
Abstract: This protocol describes the process of Phage Immunoprecipitation Sequencing (PhIP-Seq) as it is performed in the DeRisi lab. It is the basis of many published papers from our group and is used for the discovery of protein targets of antibodies. The technique uses T7 bacteriophage that display a library of tiled peptide fragments that cover a human or microbial proteome. Human sera or monoclonal antibody controls are incubated with a phage display library and antibodies are immunoprecipitated using protein A/G beads. The purified phage library is then amplified in E. coli and re-incubated with sera, typically using three rounds of enrichment. Enriched phage DNA is then sequenced to identify antibody reactivity. In this way, a proteomic assay (immunoprecipitation) is converted into a genomic assay, leveraging the significant advantages of next-generation sequencing counting statistics.
Authors: Hannah Kortbawi, Caleigh Mandell-Brehm, Brian O'Donovan, Saravazquez not provided, Madhura raghavan, Elze Rackaityte, Grace Wang, Aditi Saxena, David Yu, Joseph Derisi
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W4391127662