Generation and analyses of human synthetic antibody libraries and their application for protein microarrays
Säll, Anna LU ; Walle, Maria LU ; Wingren, Christer LU ; Müller, Susanne ; Nyman, Tomas ; Vala, Andrea ; Ohlin, Mats LU
; Borrebaeck, Carl A K
LU
and Persson, Helena
LU
(2016)
In Protein Engineering Design & Selection
29(10).
p.427-437
- 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... (More)
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.
(Less)
- author
- Säll, Anna
LU
; Walle, Maria
LU
; Wingren, Christer
LU
; Müller, Susanne
; Nyman, Tomas
; Vala, Andrea
; Ohlin, Mats
LU
; Borrebaeck, Carl A K
LU
and Persson, Helena
LU
- organization
- publishing date
- 2016-10-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- affinity proteomics, phage display technology, protein microarrays, scFv, synthetic antibody libraries
- in
- Protein Engineering Design & Selection
- volume
- 29
- issue
- 10
- pages
- 11 pages
- publisher
- Oxford University Press
- external identifiers
-
- pmid:27590051
- wos:000386205100005
- scopus:84990890044
- ISSN
- 1741-0126
- DOI
- 10.1093/protein/gzw042
- language
- English
- LU publication?
- yes
- id
- b31dd1d9-6a8f-4555-8ebe-d985d831e0b1
- date added to LUP
- 2016-11-02 09:01:39
- date last changed
- 2024-11-30 11:21:36
@article{b31dd1d9-6a8f-4555-8ebe-d985d831e0b1,
abstract = {{<p>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.</p>}},
author = {{Säll, Anna and Walle, Maria and Wingren, Christer and Müller, Susanne and Nyman, Tomas and Vala, Andrea and Ohlin, Mats and Borrebaeck, Carl A K and Persson, Helena}},
issn = {{1741-0126}},
keywords = {{affinity proteomics; phage display technology; protein microarrays; scFv; synthetic antibody libraries}},
language = {{eng}},
month = {{10}},
number = {{10}},
pages = {{427--437}},
publisher = {{Oxford University Press}},
series = {{Protein Engineering Design & Selection}},
title = {{Generation and analyses of human synthetic antibody libraries and their application for protein microarrays}},
url = {{http://dx.doi.org/10.1093/protein/gzw042}},
doi = {{10.1093/protein/gzw042}},
volume = {{29}},
year = {{2016}},
}