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Measurement and comparison of immune diversity by high-throughput sequencing

  • US 9,909,180 B2
  • Filed: 02/04/2014
  • Issued: 03/06/2018
  • Est. Priority Date: 02/04/2013
  • Status: Active Grant
First Claim
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1. A method of identifying an antigen-specific antibody lineage in a subject exposed to the antigen, comprising,sequencing nucleic acid from a biological sample obtained from the subject after exposure to the antigen, via high throughput sequencing, to determine CDR3 nucleic acid sequences, representing at least a portion of the subject'"'"'s antibody repertoire;

  • performing a linkage analysis of the CDR3 nucleic acid sequences to establish linked CDR3 sequences, wherein two CDR3 sequences are in a linkage if they have the same V and J assignment and no more than one amino acid difference in the CDR3 region;

    grouping CDR3 sequences into a plurality of lineages, wherein individual lineages of the plurality comprise (i) identical CDR3 sequences and (ii) CDR3 sequences that are associated with one another through one or more linkages; and

    comparing the CDR3 sequences in the plurality of lineages with a CDR3 sequence of an antigen-specific antibody obtained from the subject to determine the presence of the antigen-specific antibody lineage.

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