VirScan™ PhIP-Seq Antibody Profiling Service
VirScan™, an application of Bacteriophage-Display Immunoprecipitation Sequencing (PhIP-Seq), is composed of all known vertebrate viral proteomes.REQUEST INFO
VirScan enables simultaneous epitope-level autoantibody profiling against an entire virion proteome via phage display and immunoprecipitation sequencing (PhIP-Seq).
VirScan enables simultaneous epitope-level autoantibody profiling against an entire virion proteome.
The VirScan PhIP-Seq library is designed to target >68,000 annotated viral protein sequences from most known proteins from vertebrate, mosquito-borne, and tick-borne viral genomes according to the UniProt and RefSeq databases in 2017. VirScan contains an E. coli codon-optimized library consisting of >480,000 oligonucleotides encoding 62-mer amino acid peptide inserts with 14-mer amino acid adjacent overlaps.
The oligonucleotides library was synthesized on a releasable microarray, PCR amplified using appended adapter sequences, and cloned en masse into a modified T7-Select vector. This vector displays a library peptide as a fusion to the T7 phage capsid protein. The library was then packaged in vitro and expanded. The quality of library inserts was assessed by Illumina sequencing.
Screens are performed in PBS which contains phage library at an average of 105 excess clones per unique phage in the input library. The mixture is rotated overnight to allow antibodies to bind any phage-displayed peptide targets. The next day, a Protein A/G coated magnetic bead slurry is added and rotated for an additional 4 hours to capture all IgG. A set of independent Protein A/G beads-only mock-immunoprecipitation buffer-alone samples are included as negative controls. All protein A/G beads are then washed and resuspended with a Herculase II Fusion Polymerase master mix and amplified with PCR. Multiplex barcodes and sequencing adapters are then incorporated during a subsequent PCR reaction. Barcoded samples are pooled and sequenced using Illumina instruments to obtain paired-end reads of the amplified phage inserts.
A VirScan service involves case and control serum or plasma samples. These undergo a protein A/G pulldown assay, PCR amplification, and next-generation sequencing. Raw sequence data are run through a normalization and quantitation pipeline. Raw pipeline counts outputs are then provided to customers alongside normalized hit calls data for all individual 62mer peptides.
Individual PhIP-Seq studies are prepared in x96 well plates using aliquots of our phage library; each study requires x48 controls pooled plasma, known polyclonal, and protein A/G beads-only internal control samples per sequencer run (provided free-of-charge). Data are most reproducible within a single sequencer run. Sequencer runs are currently limited to 4x 96-well plates (336 experimental samples + x48 controls per run).
We will include shipping details in your quote – you must cover the cost of shipping samples to CDI Labs. Typically, after you receive your report, CDI Labs 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. Return shipping will be charged.