Carl Borrebaeck
Professor
Generation and analyses of human synthetic antibody libraries and their application for protein microarrays
Author
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