PhIP-Seq: PHAGE DISPLAY IMMUNOPRECIPITATION SEQUENCING
Overview of PhIP-Seq.
Bacteriophage Immunoprecipitation Sequencing (PhIP-Seq) is a powerful new method of multiplexed analysis that combines DNA high-throughput sequencing with next-generation proteomics. PhIP-Seq allows researchers to determine what antibodies were created by an individual’s unique history of immune exposures. PhIP-Seq is the highest-coverage antigen-specific assay yet developed, able to detect antibodies versus virtually any antigen via overlapping long peptides.
PhIP-Seq was developed at Harvard University in 2011. This work created a synthetic representation of the complete human proteome as a T7 bacteriophage display library. They built upon this work in 2015 by creating a synthetic representation of the complete human virome: the universe of known infectious viruses.
Since then, teams at UCSF and the Chan Zuckerberg Biohub have built upon and expanded the universe of antigen-specific profiling available via PhiP-Seq. PhIP-Seq has been used to perform revolutionary immune profiling on people suffering from multiple sclerosis, diabetes, rheumatoid arthritis, and more. CDI Labs was the first company to commercialize PhIP-Seq technology.
Technology overview.
PhIP-Seq: HuScan™ — Human proteome phage display.
The entire human proteome in a single antibody detection assay.
HuScan™ v2.0, an application of Bacteriophage-Display Immunoprecipitation Sequencing (PhIP-Seq), is composed of an entire normal human proteome as defined by the NCBI RefSeq database. The library contains proteins expressed as overlapping 49mer peptide tiles on the surface of bacteriophages. For an assay, an aliquot from this library is reacted with diluted patient serum or other antibody-containing fluid. Bound antibodies are immunoprecipitated with protein A/G beads, the precipitate amplified by PCR, and the sequences quantified by a next-generation sequencing and analysis pipeline that compares patient-sample IP read counts to negative controls with no antibody input (mock IPs) in the context of overall clonal frequency of individual peptides in the parent library. Output data are then created at both the peptide and whole-protein level.
PhIP-Seq: MouseScan™ — Murine proteome phage display.
The entire mouse proteome in a single antibody detection assay.
MouseScan™, an application of Bacteriophage-Display Immunoprecipitation Sequencing (PhIP-Seq), is composed of the entire normal mouse proteome (GRCm38.p5). The library contains all proteins expressed as overlapping 62mer peptide tiles on the surface of bacteriophages. For an assay, an aliquot from this library is reacted with diluted mouse serum or other antibody-containing fluid. Bound antibodies are immunoprecipitated with protein A/G beads, the precipitate amplified by PCR, and the sequences quantified by a next-generation sequencing and analysis pipeline that compares patient-sample IP read counts to negative controls with no antibody input (mock IPs) in the context of overall clonal frequency of individual peptides in the parent library. Output data are then created at both the peptide level.