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

Author:
  • Anna Säll
  • Maria Walle
  • Christer Wingren
  • Susanne Müller
  • Tomas Nyman
  • Andrea Vala
  • Mats Ohlin
  • Carl A K Borrebaeck
  • Helena Persson
Publishing year: 2016-10-01
Language: English
Pages: 427-437
Publication/Series: Protein Engineering, Design and Selection
Volume: 29
Issue: 10
Document type: Journal article
Publisher: Oxford University Press

Abstract english

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.

Keywords

  • Medical Biotechnology
  • affinity proteomics
  • phage display technology
  • protein microarrays
  • scFv
  • synthetic antibody libraries

Other

Published
  • ISSN: 1741-0126
Carl Borrebaeck
E-mail: carl [dot] borrebaeck [at] immun [dot] lth [dot] se

Professor

Department of Immunotechnology

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