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Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies
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Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies
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Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies
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Journal Article
Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies
2019
Overview
Antibody phage display has been pivotal in the quest to generate human monoclonal antibodies for biomedical and research applications. Target antigen preparation is a main bottleneck associated with the panning process. This includes production complexity, downstream purification, quality and yield. In many instances, purified antigens are preferred for panning but this may not be possible for certain difficult target antigens. Here, we describe an improved procedure of affinity selection against crude or non-purified antigen by saturation of non-binders with blocking agents to promote positive binder enrichment termed as
Yin-Yang
panning. A naïve human scFv library with kappa light chain repertoire with a library size of 10
9
was developed. The improved
Yin-Yang
biopanning process was able to enrich monoclonal antibodies specific to the MERS-CoV nucleoprotein. Three unique monoclonal antibodies were isolated in the process. The
Yin-Yang
biopanning method highlights the possibility of utilizing crude antigens for the isolation of monoclonal antibodies by phage display.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 692/4017
/ 82
/ 82/1
/ 82/80
/ 96/1
/ Antibodies, Viral - immunology
/ Antibodies, Viral - isolation & purification
/ Antigens
/ Antigens, Viral - immunology
/ Binders
/ Cell Surface Display Techniques - methods
/ Humanities and Social Sciences
/ Humans
/ Immunoglobulin kappa-Chains - immunology
/ Immunoglobulin kappa-Chains - isolation & purification
/ Middle East Respiratory Syndrome Coronavirus - immunology
/ Nucleocapsid Proteins - immunology
/ Panning
/ Science
