Journal
PROTEIN ENGINEERING DESIGN & SELECTION
Volume 29, Issue 10, Pages 427-437Publisher
OXFORD UNIV PRESS
DOI: 10.1093/protein/gzw042
Keywords
affinity proteomics; phage display technology; protein microarrays; scFv; synthetic antibody libraries
Funding
- EU FP7 AFFINOMICS: Protein Binders for Characterization of Human Proteome Function: Generation, Validation, Application [241481]
- Novo Nordisk Foundation Centre for Protein Research
- Novo Nordisk Foundation [NNF14CC0001]
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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.
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