Generation and analyses of human synthetic antibody libraries and their application for protein microarrays

Research output: Contribution to journalArticle

Abstract

Antibody-based proteomics offers distinct advantages in the analysis of complex samples for discovery and validation of biomarkers associated with disease. However, its large-scale implementation requires tools and technologies that allow development of suitable antibody or antibody fragments in a high-throughput manner. To address this we designed and constructed two human synthetic antibody fragment (scFv) libraries denoted HelL-11 and HelL-13. By the use of phage display technology, in total 466 unique scFv antibodies specific for 114 different antigens were generated. The specificities of these antibodies were analyzed in a variety of immunochemical assays and a subset was further evaluated for functionality in protein microarray applications. This high-throughput approach demonstrates the ability to rapidly generate a wealth of reagents not only for proteome research, but potentially also for diagnostics and therapeutics. In addition, this work provides a great example on how a synthetic approach can be used to optimize library designs. By having precise control of the diversity introduced into the antigen-binding sites, synthetic libraries offer increased understanding of how different diversity contributes to antibody binding reactivity and stability, thereby providing the key to future library optimization.

Details

Authors
Organisations
External organisations
  • KTH Royal Institute of Technology
  • University of Oxford
  • Karolinska Institutet
  • University of Copenhagen
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Medical Biotechnology

Keywords

  • affinity proteomics, phage display technology, protein microarrays, scFv, synthetic antibody libraries
Original languageEnglish
Pages (from-to)427-437
Number of pages11
JournalProtein Engineering, Design and Selection
Volume29
Issue number10
Publication statusPublished - 2016 Oct 1
Publication categoryResearch
Peer-reviewedYes