HuScan: PhIP-Seq Antibody Profiling

Human proteome epitope-level autoantibody profiling service via T7 phage display and immunonprecipitation sequencing

HuScan: PhIP-Seq Antibody Profiling

Human proteome epitope-level autoantibody profiling service via T7 phage display and immunonprecipitation sequencing

On this page you can order HuScan, a human autoantibody discovery service by CDI Labs.

Our new HuScan™ technology provides epitope-level antibody discovery versus the entire human proteome.

ANTYGENHuScan™ service, co-developed by CDI’s Scientific Advisory Board Member Ben Larman, allows us to analyze patient samples (serum, plasma, CSF, etc.) for the presence of autoantibodies against the full UniProt human proteome in a single-well assay with quantitative sequencing readouts. This product is available as a service to both academic and industry customers.

Diagram of VirScan PhIP-Seq Antygen Service Product by CDI Labs

Download an overview of our HuScan™ service.

CDI Antygen™ HuScan™is provided as a complete analysis service.

Understanding Your HuScan and VirScan Results

HuScan™ technical details.

The original HuScan PhIP-Seq library was created by downloading viral protein sequences from UniProt, using software to tile overlapping sequences, tuning tiled sequences for translation by the phages and their bacterial hosts, and then ordering the sequences as synthetic oligonucleotides from a vendor as described (Larman, et al, 2011). This oligonucleotide library was PCR-amplified with adaptors for cloning and inserted into a T7 phage display vector that was expanded in E. Coli. The expanded T7 phage library quality is confirmed by sequencing to have >90% of the library within one log of overall average clonal frequency. An aliquot from this library is then 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. A more detailed description of this process is available (Mohan, et al, 2018).

HuScan™ service details & data deliverables.

A typical HuScan™ service involves case and control serum or plasma samples. These are heat deactivated and undergo a protein A/G pulldown assay, PCR amplification, and next-generation sequencing. Raw sequence data are run through a normalization and quantitation pipeline as previously described (Mohan et al., 2018).  Raw pipeline outputs are then provided to customers as normalized read counts data tables for both whole proteins and individual 90mer peptides.

HuScan™ service sample requirements.

Human serum or plasma: Send two 20μL aliquots per sample.

Human (CSF) cerebrospinal fluid: Send two 250μL aliquots per sample.

Ten sample minimum.

Note: VirScan™ and HuScan™ can be run together in a single-well assay at reduced cost.

Sample shipping & sample return.

We will include shipping details in your quote – you must cover the cost of shipping samples to CDI. Typically, after you receive your report, CDI keeps the remaining samples for two months and then disposes of them. When shipping to us, please let us know if you want the remaining samples returned after the study is complete. We will charge for return shipping.

Contact us to learn more.

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