Isolation and structure–functional characterization of phage display library-derived mimotopes of noxiustoxin, a neurotoxin of the scorpion Centruroides noxius Hoffmann

Viral infection and pathogenesis is mediated by host protein—viral protein complexes that are important targets for therapeutic intervention as they are potentially less prone to development of drug resistance. We have identified human, recombinant antibodies (Fabs) from a phage display library that bind to three HIV-host complexes. We used these Fabs to 1) stabilize the complexes for structural studies; and 2) facilitate characterization of the function of these complexes. Specifically, we generated recombinant Fabs to Vif-CBF-β-ELOB-ELOC (VCBC); ESCRT-I complex and AP2-complex. For each complex we measured binding affinities with KD values of Fabs ranging from 12–419 nM and performed negative stain electron microscopy (nsEM) to obtain low-resolution structures of the HIV-Fab complexes. Select Fabs were converted to scFvs to allow them to fold intracellularly and perturb HIV-host protein complex assembly without affecting other pathways. To identify these recombinant Fabs, we developed a rapid screening pipeline that uses quantitative ELISAs and nsEM to establish whether the Fabs have overlapping or independent epitopes. This pipeline approach is generally applicable to other particularly challenging antigens that are refractory to immunization strategies for antibody generation including multi-protein complexes providing specific, reproducible, and renewable antibody reagents for research and clinical applications. The curated antibodies described here are available to the scientific community for further structural and functional studies on these critical HIV host-factor proteins.

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Genetic and functional characterization of human pemphigus vulgaris monoclonal autoantibodies isolated by phage display

Journal of Clinical Investigation ◽

10.1172/jci24185 ◽

2005 ◽

Vol 115 (4) ◽

pp. 888-899 ◽

Cited By ~ 143

Author(s):

Aimee S. Payne ◽

Ken Ishii ◽

Stephen Kacir ◽

Chenyan Lin ◽

Hong Li ◽

...

Keyword(s):

Phage Display ◽

Functional Characterization ◽

Pemphigus Vulgaris

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Isolation of and Characterization of Neutralizing Antibodies to Covid-19 from a Large Human Naïve scFv Phage Display Library

10.1101/2020.05.19.104281 ◽

2020 ◽

Cited By ~ 6

Author(s):

Andy Q. Yuan ◽

Likun Zhao ◽

Lili Bai ◽

Qingwu Meng ◽

Zhenguo Wen ◽

...

Keyword(s):

Phage Display ◽

Neutralizing Antibodies ◽

Epitope Mapping ◽

Neutralization Assay ◽

Phage Display Library ◽

Antibody Library ◽

Antibody Phage ◽

Antibody Phage Display ◽

Human Receptor

AbstractSARS-CoV-2 (Covid-19) has caused currently ongoing global plague and imposed great challenges to health managing systems all over the world, with millions of infections and hundreds of thousands of deaths. In addition to racing to develop vaccines, neutralizing antibodies (nAbs) to this virus have been extensively sought and are expected to provide another prevention and therapy tool against this frantic pandemic. To offer fast isolation and shortened early development, a large human naïve phage display antibody library, was built and used to screen specific nAbs to the receptor-binding domain, RBD, the key for Covid-19 virus entry through a human receptor, ACE2. The obtained RBD-specific antibodies were characterized by epitope mapping, FACS and neutralization assay. Some of the antibodies demonstrated spike-neutralizing property and ACE2-competitiveness. Our work proved that RBD-specific neutralizing binders from human naïve antibody phage display library are promising candidates to for further Covid-19 therapeutics development.

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