Generation of MANAbodies specific to HLA-restricted epitopes encoded by somatically mutated genes

Andrew D. Skora, Jacqueline Douglass, Michael S. Hwang, Ada J. Tam, Richard L. Blosser, Sandra B. Gabelli, Jianhong Cao, Luis A. Diaz, Nickolas Papadopoulos, Kenneth W. Kinzler, Bert Vogelstein, Shibin Zhou

Research output: Contribution to journalArticle

Abstract

Mutant epitopes encoded by cancer genes are virtually always located in the interior of cells, making them invisible to conventional antibodies. We here describe an approach to identify single-chain variable fragments (scFvs) specific for mutant peptides presented on the cell surface by HLA molecules. We demonstrate that these scFvs can be successfully converted to full-length antibodies, termed MANAbodies, targeting "Mutation-Associated Neo-Antigens" bound to HLA. A phage display library representing a highly diverse array of single-chain variable fragment sequences was first designed and constructed. A competitive selection protocol was then used to identify clones specific for mutant peptides bound to predefined HLA types. In this way, we obtained two scFvs, one specific for a peptide encoded by a common KRAS mutant and the other by a common epidermal growth factor receptor (EGFR) mutant. The scFvs bound to these peptides only when the peptides were complexed with HLA-A2 (KRAS peptide) or HLA-A3 (EGFR peptide). We converted one scFv to a fulllength antibody (MANAbody) and demonstrate that the MANAbody specifically reacts with mutant peptide-HLA complex even when the peptide differs by only one amino acid from the normal, WT form.

Original languageEnglish (US)
Pages (from-to)9967-9972
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number32
DOIs
StatePublished - Aug 11 2015

Keywords

  • Antibody engineering
  • Immunotherapy
  • Oncogene
  • Personalized
  • Therapy

ASJC Scopus subject areas

  • General

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