title: Generation and analyses of human synthetic antibody libraries and their application for protein microarrays.
abstract: 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.
authors: Department of Immunotechnology, Lund University, Medicon Village (Bldg 406), 223 81 Lund, Sweden.
authors: Anna
authors: A
authors: Säll
authors: Department of Immunotechnology, Lund University, Medicon Village (Bldg 406), 223 81 Lund, Sweden.
authors: Maria
authors: M
authors: Walle
authors: Department of Immunotechnology, Lund University, Medicon Village (Bldg 406), 223 81 Lund, Sweden.
authors: Christer
authors: C
authors: Wingren
authors: The Structural Genomics Consortium, University of Oxford, Roosevelt Drive, OxfordOX3 7FZ, UK.
authors: Susanne
authors: S
authors: Müller
authors: Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Scheeles väg 2, 171 77 Stockholm, Sweden.
authors: Tomas
authors: T
authors: Nyman
authors: The Novo Nordisk Foundation Centre for Protein Research, Protein Structure and Function Program, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2000 Copenhagen, Denmark.
authors: Andrea
authors: A
authors: Vala
authors: Department of Immunotechnology, Lund University, Medicon Village (Bldg 406), 223 81 Lund, Sweden.
authors: Mats
authors: M
authors: Ohlin
authors: Department of Immunotechnology, Lund University, Medicon Village (Bldg 406), 223 81 Lund, Sweden.
authors: Carl A K
authors: CAK
authors: Borrebaeck
authors: Department of Immunotechnology, Lund University, Medicon Village (Bldg 406), 223 81 Lund, Sweden.
authors: Helena
authors: H
authors: Persson
mesh_terms: D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D006801:Humans; D019151:Peptide Library; D040081:Protein Array Analysis / Q000379:methods*; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*
chemical_list: D000941:Antigens; D019151:Peptide Library; D057127:Single-Chain Antibodies
keywords: affinity proteomics; phage display technology; protein microarrays; scFv; synthetic antibody libraries