Generation and analyses of human synthetic antibody libraries and their application for protein microarrays
| Autor: | Maria Walle, Helena Persson, Tomas Nyman, Christer Wingren, Mats Ohlin, Anna Säll, Andrea Vala, Carl A.K. Borrebaeck, Susanne Müller |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Phage display Protein Array Analysis Bioengineering Computational biology Biology Proteomics Biochemistry Molecular biology Synthetic antibody 03 medical and health sciences 030104 developmental biology Antibody Specificity Peptide Library Proteome Protein microarray Humans Antigens Peptide library Molecular Biology Single-Chain Antibodies Biotechnology |
| Zdroj: | Protein Engineering Design and Selection. 29:427-437 |
| ISSN: | 1741-0134 1741-0126 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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