HuScan™, VirScan™, & XenScan: PHAGE DISPLAY IMMUNOPRECIPITATION SEQUENCING

Overview of HuScan™, VirScan™, & XenScan™ 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. Our PhIP-Seq assay is the highest-coverage antigen-specific assay ever developed, able to detect antibodies versus every known human protein or virus as overlapping long peptides.

PhIP-Seq was developed at Harvard University by CDI Scientific Advisory Board member Ben Larman, Steve Elledge, and colleagues in 2011. This work created HuScan™, a synthetic representation of the complete human proteome as a T7 bacteriophage display library. They built upon this work in 2015, by creating VirScan™, a synthetic representation of the complete human virome: the universe of known infectious viruses.

Since then, PhIP-Seq has been used to perform revolutionary immune profiling on people suffering from measles, multiple sclerosis, diabetes, rheumatoid arthritis, and more. By sequencing millions of DNA strands at once, CDI’s PhIP-Seq is a powerful technology that helps scientists determine the immune system’s role in complex disease non-invasively, quickly, and at a low cost.

Creation of the XenScan™ Immune Epitope Database (IEDB) xenoantigen profiling assay was first reported alongside an updated VirScan™ library.  This library contains every full-length UniProt protein with an IEDB-curated antigen that does not originate from humans, viruses, or malaria (plasmodium).

Technology overview.

PhIP-Seq: HuScan™ — Human proteome phage display.

The entire human proteome in a single antibody detection assay.

HuScan™, an application of Bacteriophage-Display Immunoprecipitation Sequencing (PhIP-Seq), is composed of the entire normal human proteome as defined by the UniProt database. The library contains all proteins expressed as overlapping 90mer 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.

Learn more about our ANTYGEN™ HuScan™ analysis services standalone or as part of a Grand Seromics assay:

Learn more about our ANTYGEN™ HuScan™ analysis services.

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PhIP-Seq: VirScan™ — Human virome phage display.

Most known human viruses in a single antibody detection assay.

VirScan™, an application of Bacteriophage-Display Immunoprecipitation Sequencing (PhIP-Seq), is composed of >1000 individual strains across 206 unique viral species with known human tropism present in the UniProt database. The library contains all proteins from these viruses expressed as overlapping 56mer 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.

Learn more about our ANTYGEN™ VirScan™ analysis services standalone or as part of a Grand Seromics assay:

Learn more about our ANTYGEN™ VirScan™ analysis services.

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VirScan references.

PhIP-Seq: XenScan™ — IEDB xenoantigen phage display.

The most important xenoantigens in a single antibody detection assay.

XenScan™, an application of Bacteriophage-Display Immunoprecipitation Sequencing (PhIP-Seq), is composed of 604 individual Immune Epitope Database (IEDB) xenoantigens across 213 unique species. The library contains full-length proteins encompassing all IEDB-curated antigens that does not originate from humans, viruses, or malaria (plasmodium) expressed as overlapping 56mer 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.

Learn more about our ANTYGEN™ XenScan™ analysis services.

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XenScan references.