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Carl B

Carl Borrebaeck

Professor

Carl B

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

Summary, in 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.

Department/s

  • Department of Immunotechnology
  • BioCARE: Biomarkers in Cancer Medicine improving Health Care, Education and Innovation

Publishing year

2016-10-01

Language

English

Pages

427-437

Publication/Series

Protein Engineering Design & Selection

Volume

29

Issue

10

Document type

Journal article

Publisher

Oxford University Press

Topic

  • Medical Biotechnology

Keywords

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

Status

Published

ISBN/ISSN/Other

  • ISSN: 1741-0126