Combinatorial libraries against libraries for selecting neoepitope activation-specific antibodies

Xuebo Hu, Sungkwon Kang, Craig Lefort, Minsoo Kim, Moonsoo M. Jin

Research output: Contribution to journalArticlepeer-review

21 Scopus citations


A systematic approach to the discovery of conformation-specific antibodies or those that recognize activation-induced neoepitopes in signaling molecules and enzymes will be a powerful tool in developing antibodies for basic science and therapy. Here, we report the isolation of antibody antagonists that preferentially bind activated integrin Mac-1 (αMβ 2) and are potent in blocking neutrophil adhesion and migration. A novel strategy was developed for this task, consisting of yeast surface display of Mac-1 inserted (I) domain library, directed evolution to isolate active mutants of the I domain, and screening of phage display of human antibody library against the active I domain in yeast. Enriched antibody library was then introduced into yeast surface two-hybrid system for final quantitative selection of antibodies from monomeric antigen-antibody interaction. This led to highly efficient isolation of intermediate to high affinity antibodies, which preferentially reacted with the active I domain, antagonized the I domain binding to intercellular adhesion molecule (ICAM)-1, complement C3 fragment iC3b, and fibronectin, and potently inhibited neutrophil migration on fibrinogen. The strategy demonstrated herein can be broadly applicable to developing antibodies against modular domains that switch between inactive and active conformations, particularly toward the discovery of antibody antagonists in therapeutic and diagnostic applications.

Original languageEnglish (US)
Pages (from-to)6252-6257
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number14
StatePublished - Apr 6 2010


  • Antibody antagonist
  • Mac-1 integrin
  • Phage display
  • Yeast display

ASJC Scopus subject areas

  • General


Dive into the research topics of 'Combinatorial libraries against libraries for selecting neoepitope activation-specific antibodies'. Together they form a unique fingerprint.

Cite this