Archer® Immunoverse™ kits are targeted NGS assays to characterize the human immune repertoire from RNA input. Powered by Anchored Multiplex PCR (AMP™), the lyophilized kits uniquely tag and amplify V(D)J rearrangements for sequencing on Illumina® platforms. Sequenced libraries are analyzed using the Immune Repertoire pipeline in Archer Analysis—a powerful and transparent tool for clonotype identification and frequency reporting.
Immunoverse TCR kits amplify T cell receptor beta/gamma or alpha/delta chains for sequencing and analysis across a wide variety of clonality and diversity applications. With targeted primers within 100 bp of the end of the CDR3 sequence of interest, the beta/gamma kit is designed for maximal sensitivity in FFPE material to profile tumor infiltrating lymphocyte (TILs). Sample inputs range from 25ng to 2μg and can be sequenced at any depth on 2x150 bp Illumina kits.
Immunoverse BCR kits amplify B cell receptor heavy (IGH) and light (kappa/lambda) chains for sequencing and analysis. Primers are designed to identify isotypes and subclasses in the Fc region and somatic hypermutations within the variable regions. Sample inputs range from 25ng and 6μg and can be sequenced at any depth on 2x150 bp or 2x300 bp Illumina kits.
Archer Analysis, a powerful and transparent tool for immune repertoire analysis, providing extensive clone and clonotype information in custom-filtered tables, dynamic visualizations, and full data exports to answer advanced research questions. Archer analysis is available for private cloud and local installation behind your firewall.
Jurkat cell line dilution in PBL background down to 10-6 from 400ng input shows strong linearity and detection of jurkat sequences at expected frequencies across replicates
Libraries were prepped using the Immunoverse TCR beta/gamma kit and sequenced on an Illumina® NextSeq® using a 300-cycle kit. Libraries were normalized to 0.5M reads and 2.5M reads (dilution 6) and analyzed with Archer Analysis.
Highly reproducible clonotype frequency reporting of top 10 clones between replicates in jurkat dilution into PBL background
Libraries were prepped from 400ng input using the Immunoverse TCR beta/gamma kit and sequenced on an Illumina® MiSeq® using a 600-cycle kit. Libraries were normalized to 0.6M reads and analyzed with Archer Analysis.
Strong linearity and reproducibility is shown in dilution series across two different laboratories
Ten-fold serial dilutions of Jurkat cell line mRNA into PBL background we performed in duplicates in two labs. Libraries were sequenced on Illumina® NextSeq® with 2x150bp reads and normalized to 0.5M reads and analyzed using archer analysis.
Archer analysis reports somatic hypermutation in most frequent clonotypes with >2% deviation in reference V-region homology.
400ng of a Chronic Lymphocytic Leukemia (CLL) sample was prepped with the Immunoverse BCR IGH kit. Libraries were sequenced on the Illumina MiSeq® v3 (600 cycle) kit and normalized to 360,000 reads. Reads were Deduplicated and error corrected using Archer Analysis, clonal frequencies and somatic hypermutation was reported for the top 5 clones according to the ERIC recommendations.
The Immunoverse assays utilize AMP chemistry for open-ended amplification from molecular barcoded (MBC) adapters, eliminating the need for complicated opposing primer-based amplification schemes and uniformity assumptions. This unbiased approach towards VDJ recombination can yield more than 100,000 clonotypes in a single reaction, without the fear of PCR bias. The figure above shows a simplified schematic of AMP-enabled V(D)J enrichment.
Archer Analysis utilizes the AMP-specific molecular barcode adapters for deduplication and PCR sequencing error correction to provide a true measure of sample complexity. Additionally, unique molecule identification enables highly quantitative analysis of clonotype frequencies.
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