Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation
Antibodies leverage on their unique architecture to bind with an array of antigens. The strength of interaction has a direct relation to the affinity of the antibodies towards the antigen. <i>In vivo</i> affinity maturation is performed through multiple rounds of somatic hypermutation an...
Main Authors: | , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2019-04-01
|
Series: | International Journal of Molecular Sciences |
Subjects: | |
Online Access: | https://www.mdpi.com/1422-0067/20/8/1861 |
id |
doaj-89fce58b220d40c7b6383257c6aa9932 |
---|---|
record_format |
Article |
spelling |
doaj-89fce58b220d40c7b6383257c6aa99322020-11-24T21:49:09ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-04-01208186110.3390/ijms20081861ijms20081861Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity MaturationChia Chiu Lim0Yee Siew Choong1Theam Soon Lim2Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang 11800, MalaysiaInstitute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang 11800, MalaysiaInstitute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang 11800, MalaysiaAntibodies leverage on their unique architecture to bind with an array of antigens. The strength of interaction has a direct relation to the affinity of the antibodies towards the antigen. <i>In vivo</i> affinity maturation is performed through multiple rounds of somatic hypermutation and selection in the germinal centre. This unique process involves intricate sequence rearrangements at the gene level via molecular mechanisms. The emergence of <i>in vitro</i> display technologies, mainly phage display and recombinant DNA technology, has helped revolutionize the way antibody improvements are being carried out in the laboratory. The adaptation of molecular approaches <i>in vitro</i> to replicate the <i>in vivo</i> processes has allowed for improvements in the way recombinant antibodies are designed and tuned. Combinatorial libraries, consisting of a myriad of possible antibodies, are capable of replicating the diversity of the natural human antibody repertoire. The isolation of target-specific antibodies with specific affinity characteristics can also be accomplished through modification of stringent protocols. Despite the ability to screen and select for high-affinity binders, some ‘fine tuning’ may be required to enhance antibody binding in terms of its affinity. This review will provide a brief account of phage display technology used for antibody generation followed by a summary of different combinatorial library characteristics. The review will focus on available strategies, which include molecular approaches, next generation sequencing, and <i>in silico</i> approaches used for antibody affinity maturation in both therapeutic and diagnostic applications.https://www.mdpi.com/1422-0067/20/8/1861human monoclonal antibodiesphage displaycombinatorial librariesaffinity maturation |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chia Chiu Lim Yee Siew Choong Theam Soon Lim |
spellingShingle |
Chia Chiu Lim Yee Siew Choong Theam Soon Lim Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation International Journal of Molecular Sciences human monoclonal antibodies phage display combinatorial libraries affinity maturation |
author_facet |
Chia Chiu Lim Yee Siew Choong Theam Soon Lim |
author_sort |
Chia Chiu Lim |
title |
Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation |
title_short |
Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation |
title_full |
Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation |
title_fullStr |
Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation |
title_full_unstemmed |
Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation |
title_sort |
cognizance of molecular methods for the generation of mutagenic phage display antibody libraries for affinity maturation |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2019-04-01 |
description |
Antibodies leverage on their unique architecture to bind with an array of antigens. The strength of interaction has a direct relation to the affinity of the antibodies towards the antigen. <i>In vivo</i> affinity maturation is performed through multiple rounds of somatic hypermutation and selection in the germinal centre. This unique process involves intricate sequence rearrangements at the gene level via molecular mechanisms. The emergence of <i>in vitro</i> display technologies, mainly phage display and recombinant DNA technology, has helped revolutionize the way antibody improvements are being carried out in the laboratory. The adaptation of molecular approaches <i>in vitro</i> to replicate the <i>in vivo</i> processes has allowed for improvements in the way recombinant antibodies are designed and tuned. Combinatorial libraries, consisting of a myriad of possible antibodies, are capable of replicating the diversity of the natural human antibody repertoire. The isolation of target-specific antibodies with specific affinity characteristics can also be accomplished through modification of stringent protocols. Despite the ability to screen and select for high-affinity binders, some ‘fine tuning’ may be required to enhance antibody binding in terms of its affinity. This review will provide a brief account of phage display technology used for antibody generation followed by a summary of different combinatorial library characteristics. The review will focus on available strategies, which include molecular approaches, next generation sequencing, and <i>in silico</i> approaches used for antibody affinity maturation in both therapeutic and diagnostic applications. |
topic |
human monoclonal antibodies phage display combinatorial libraries affinity maturation |
url |
https://www.mdpi.com/1422-0067/20/8/1861 |
work_keys_str_mv |
AT chiachiulim cognizanceofmolecularmethodsforthegenerationofmutagenicphagedisplayantibodylibrariesforaffinitymaturation AT yeesiewchoong cognizanceofmolecularmethodsforthegenerationofmutagenicphagedisplayantibodylibrariesforaffinitymaturation AT theamsoonlim cognizanceofmolecularmethodsforthegenerationofmutagenicphagedisplayantibodylibrariesforaffinitymaturation |
_version_ |
1725889190366806016 |