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

Research output: Contribution to journalJournal articleResearchpeer-review

  • Anna Säll
  • Maria Walle
  • Christer Wingren
  • Susanne Müller
  • Tomas Nyman
  • Andrea Lages Lino Vala
  • Mats Ohlin
  • Carl A K Borrebaeck
  • Helena Persson

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.

Original languageEnglish
JournalProtein Engineering Design and Selection (Print)
Volume29
Issue number10
Pages (from-to)427-37
Number of pages11
ISSN1741-0126
DOIs
Publication statusPublished - Oct 2016

    Research areas

  • Journal Article

ID: 166506188