Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries
Phage display is a powerful technique for drug discovery in biomedical research in particular for antibody libraries. But, several technical challenges are associated with the selection process. For instance, during the panning step, the successful elution of the phages bound to the antigen is criti...
Main Authors: | , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2019-02-01
|
Series: | Methods and Protocols |
Subjects: | |
Online Access: | https://www.mdpi.com/2409-9279/2/1/17 |
id |
doaj-289f6e17d56b4268a14d29ca9b9b5da2 |
---|---|
record_format |
Article |
spelling |
doaj-289f6e17d56b4268a14d29ca9b9b5da22020-11-25T01:13:40ZengMDPI AGMethods and Protocols2409-92792019-02-01211710.3390/mps2010017mps2010017Efficient Construction and Effective Screening of Synthetic Domain Antibody LibrariesArghavan Solemani Zadeh0Alissa Grässer1Heiko Dinter2Maximilian Hermes3Katharina Schindowski4Institute for Applied Biotechnology, Biberach University of Applied Science, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute for Applied Biotechnology, Biberach University of Applied Science, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute for Applied Biotechnology, Biberach University of Applied Science, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute for Applied Biotechnology, Biberach University of Applied Science, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyInstitute for Applied Biotechnology, Biberach University of Applied Science, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, GermanyPhage display is a powerful technique for drug discovery in biomedical research in particular for antibody libraries. But, several technical challenges are associated with the selection process. For instance, during the panning step, the successful elution of the phages bound to the antigen is critical in order to avoid losing the most promising binders. Here, we present an efficient protocol to establish, screen and select synthetic libraries of domain antibodies using phage display. We do not only present suitable solutions to the above-mentioned challenges to improve elution by 50-fold, but we also present a step by step in-depth protocol with miniaturized volumes and optimized procedures to save material, costs and time for a successful phage display with domain antibodies. Hence, this protocol improves the selection process for an efficient handling process. The here presented library is based on the variable domain (vNAR) of the naturally occurring novel antibody receptor (IgNAR) from cartilage fishes. Diversity was introduced in the Complementarity-Determining Region 3 (CDR3) of the antigen-binding site with different composition and length.https://www.mdpi.com/2409-9279/2/1/17display technologyantibody engineeringsynthetic antibody libraryshark antibodyvNARphage displaypanning |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Arghavan Solemani Zadeh Alissa Grässer Heiko Dinter Maximilian Hermes Katharina Schindowski |
spellingShingle |
Arghavan Solemani Zadeh Alissa Grässer Heiko Dinter Maximilian Hermes Katharina Schindowski Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries Methods and Protocols display technology antibody engineering synthetic antibody library shark antibody vNAR phage display panning |
author_facet |
Arghavan Solemani Zadeh Alissa Grässer Heiko Dinter Maximilian Hermes Katharina Schindowski |
author_sort |
Arghavan Solemani Zadeh |
title |
Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries |
title_short |
Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries |
title_full |
Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries |
title_fullStr |
Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries |
title_full_unstemmed |
Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries |
title_sort |
efficient construction and effective screening of synthetic domain antibody libraries |
publisher |
MDPI AG |
series |
Methods and Protocols |
issn |
2409-9279 |
publishDate |
2019-02-01 |
description |
Phage display is a powerful technique for drug discovery in biomedical research in particular for antibody libraries. But, several technical challenges are associated with the selection process. For instance, during the panning step, the successful elution of the phages bound to the antigen is critical in order to avoid losing the most promising binders. Here, we present an efficient protocol to establish, screen and select synthetic libraries of domain antibodies using phage display. We do not only present suitable solutions to the above-mentioned challenges to improve elution by 50-fold, but we also present a step by step in-depth protocol with miniaturized volumes and optimized procedures to save material, costs and time for a successful phage display with domain antibodies. Hence, this protocol improves the selection process for an efficient handling process. The here presented library is based on the variable domain (vNAR) of the naturally occurring novel antibody receptor (IgNAR) from cartilage fishes. Diversity was introduced in the Complementarity-Determining Region 3 (CDR3) of the antigen-binding site with different composition and length. |
topic |
display technology antibody engineering synthetic antibody library shark antibody vNAR phage display panning |
url |
https://www.mdpi.com/2409-9279/2/1/17 |
work_keys_str_mv |
AT arghavansolemanizadeh efficientconstructionandeffectivescreeningofsyntheticdomainantibodylibraries AT alissagrasser efficientconstructionandeffectivescreeningofsyntheticdomainantibodylibraries AT heikodinter efficientconstructionandeffectivescreeningofsyntheticdomainantibodylibraries AT maximilianhermes efficientconstructionandeffectivescreeningofsyntheticdomainantibodylibraries AT katharinaschindowski efficientconstructionandeffectivescreeningofsyntheticdomainantibodylibraries |
_version_ |
1725160727082172416 |