Generation and analyses of human synthetic antibody libraries and their application for protein microarrays

TY - JOUR

T1 - Generation and analyses of human synthetic antibody libraries and their application for protein microarrays

AU - Säll, Anna

AU - Walle, Maria

AU - Wingren, Christer

AU - Müller, Susanne

AU - Nyman, Tomas

AU - Lages Lino Vala, Andrea

AU - Ohlin, Mats

AU - Borrebaeck, Carl A K

AU - Persson, Helena

N1 - © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: [email protected].

PY - 2016/10/1

Y1 - 2016/10/1

N2 - 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.

AB - 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.

KW - Journal Article

U2 - 10.1093/protein/gzw042

DO - 10.1093/protein/gzw042

M3 - Journal article

C2 - 27590051

SN - 1741-0126

VL - 29

SP - 427

EP - 437

JO - Protein Engineering, Design and Selection

JF - Protein Engineering, Design and Selection

IS - 10

ER -