scholarly journals Colony Assay for Antibody Library Screening: Outlook and Comparison to Display Screening

2018 ◽  
Author(s):  
Mieko Kato ◽  
Yoshiro Hanyu
Keyword(s):  
Library Screening ◽  
Antibody Library ◽  
Colony Assay

scholarly journals A general strategy for antibody library screening via conversion of transient target binding into permanent reporter deposition

2014 ◽  
Vol 27 (2) ◽  
pp. 41-47 ◽  
Author(s):  
A. Maass ◽  
T. Heiseler ◽  
F. Maass ◽  
J. Fritz ◽  
T. Hofmeyer ◽  
...  
Keyword(s):  
General Strategy ◽  
Library Screening ◽  
Antibody Library ◽  
Target Binding

scholarly journals Screening Antibody Libraries with Colony Assay Using scFv-Alkaline Phosphatase Fusion Proteins

Molecules ◽  
2020 ◽  
Vol 25 (12) ◽  
pp. 2905
Author(s):  
Yoshiro Hanyu ◽  
Mieko Kato
Keyword(s):  
Alkaline Phosphatase ◽  
Fusion Proteins ◽  
Direct Detection ◽  
Screening Methods ◽  
Single Chain Variable Fragment ◽  
Antibody Library ◽  
Single Chain ◽  
Library Size ◽  
Colony Assay ◽  
Antibody Libraries

Screening antibody libraries is an important step in establishing recombinant monoclonal antibodies. The colony assay can identify positive clones without almost any false-positives; however, its antibody library is smaller than those used in other recombinant screening methods such as phage display. Thus, to improve the efficiency of colony assays, it is necessary to increase library size per screening. Here, we report developing a colony assay with single-chain variable fragment (scFv) fused to the N-terminus of bacterial alkaline phosphatase (scFv-PhoA). The scFv-PhoA library was constructed in an expression vector specifically designed for this study. Use of this library allowed the successful and direct detection of positive clones exhibiting PhoA activity, without the need for a secondary antibody. Colony assay screening with scFv-PhoA is simple, rapid, offers a higher success rate than previous methods based on scFv libraries, and—most importantly—it enables high-throughput procedures.

Antibody Library Screening in Through-Hole Microarrays

2001 ◽  
pp. 95-96
Author(s):  
Tanya S. Kanigan ◽  
Colin J. H. Brenan ◽  
Tara Heitner ◽  
Robert Hess ◽  
Arrin Katz ◽  
...  
Keyword(s):  
Library Screening ◽  
Antibody Library ◽  
Through Hole

scholarly journals Novel human neutralizing mAbs specific for Spike-RBD of SARS-CoV-2

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Margherita Passariello ◽  
Chiara Gentile ◽  
Veronica Ferrucci ◽  
Emanuele Sasso ◽  
Cinzia Vetrei ◽  
...  
Keyword(s):  
Cell Entry ◽  
The Novel ◽  
Antibody Library ◽  
Native Conformation ◽  
Innovative Strategy ◽  
Effector Functions ◽  
Igg1 Mabs ◽  
Therapeutic Tools ◽  
Nanomolar Range ◽  
Potent Effector

AbstractAmong the therapies against the pandemic SARS-CoV-2 virus, monoclonal Antibodies (mAbs) targeting the Spike glycoprotein represent good candidates to interfere in the Spike/ACE2 interaction, preventing virus cell entry. Since anti-spike mAbs, used individually, might be unable to block the virus entry in the case of resistant mutations, we designed an innovative strategy for the isolation of multiple novel human scFvs specific for the binding domain (RBD) of Spike. By panning a large phage display antibody library on immobilized RBD, we obtained specific binders by eluting with ACE2 in order to identify those scFvs recognizing the epitope of Spike interacting with its receptor. We converted the novel scFvs into full size IgG4, differently from the previously isolated IgG1 mAbs, to avoid unwanted potential side effects of IgG1 potent effector functions on immune system. The novel antibodies specifically bind to RBD in a nanomolar range and interfere in the interaction of Spike with ACE2 receptor, either used as purified protein or when expressed on cells in its native conformation. Furthermore, some of them have neutralizing activity for virus infection in cell cultures by using two different SARS-CoV-2 isolates including the highly contagious VOC 202012/01 variant and could become useful therapeutic tools to fight against the SARS-CoV-2 virus.

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