×

Adaptive immunity profiling and methods for generation of monoclonal antibodies

  • US 10,323,276 B2
  • Filed: 02/06/2014
  • Issued: 06/18/2019
  • Est. Priority Date: 01/15/2009
  • Status: Active Grant
First Claim
Patent Images

1. A method for generating cDNA sequence libraries useful in producing monoclonal antibodies, the method comprising:

  • generating a pre-immunized cDNA sequence library comprising cDNA sequences encoding proteins generated from a region subject to genomic rearrangement, wherein the region is an Ig heavy chain or light chain variable domain in a B-cell, or a T-cell alpha or beta variable domain in a T-cell, wherein the cDNA sequences are obtained from a first sample comprising lymphocytes of a host subject that has not been immunized with an antigen, wherein generating the pre-immunized cDNA sequence library comprises amplifying the cDNA sequences and sequencing the amplified cDNA sequences;

    generating an immunized cDNA sequence library comprising cDNA sequences encoding proteins generated from a region subject to genomic rearrangement, wherein the region is an Ig heavy chain or light chain variable domain in a B-cell, or a T-cell alpha or beta variable domain in a T-cell, wherein the cDNA sequences are obtained from a second sample comprising lymphocytes of said host subject after being immunized with said antigen, wherein generating the immunized cDNA sequence library comprises amplifying the cDNA sequences and sequencing the amplified cDNA sequences;

    analyzing the frequencies of occurrence of each of said pre-immunized and immunized set of cDNA sequences; and

    identifying candidate cDNA sequences useful in producing monoclonal antibodies from said pre-immunized and immunized libraries of cDNA sequences by comparing the frequency of occurrence in each library of cDNA sequences, wherein each candidate cDNA sequence is one which has a significantly higher frequency of occurrence in the immunized sample as compared to the frequency of occurrence in the pre-immunized sample.

View all claims
  • 3 Assignments
Timeline View
Assignment View
    ×
    ×