Simple Paired Heavy- And Light-Chain Antibody Repertoire Sequencing Using Endoplasmic Reticulum Microsomes

Genome Med. 2018 Apr 27;10(1):34. doi: 10.1186/s13073-018-0542-5.

Abstract

Existing methods for paired antibody heavy- and light-chain repertoire sequencing rely on specialized equipment and are limited by their commercial availability and high costs. Here, we report a novel simple and cost-effective emulsion-based single-cell paired antibody repertoire sequencing method that employs only basic laboratory equipment. We performed a proof-of-concept using mixed mouse hybridoma cells and we also showed that our method can be used for discovery of novel antigen-specific monoclonal antibodies by sequencing human CD19+ B cell IgM and IgG repertoires isolated from peripheral whole blood before and seven days after Td (Tetanus toxoid/Diphtheria toxoid) booster immunization. We anticipate broad applicability of our method for providing insights into adaptive immune responses associated with various diseases, vaccinations, and cancer immunotherapies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies / chemistry
  • Antibodies / metabolism*
  • Antibodies, Monoclonal / metabolism
  • B-Lymphocytes / metabolism
  • Endoplasmic Reticulum / metabolism*
  • HEK293 Cells
  • Humans
  • Immunoglobulin Heavy Chains / metabolism
  • Immunoglobulin Light Chains / metabolism
  • Immunoglobulin Variable Region / genetics
  • Mice
  • Microsomes / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sequence Analysis, Protein

Substances

  • Antibodies
  • Antibodies, Monoclonal
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Light Chains
  • Immunoglobulin Variable Region
  • RNA, Messenger