scholarly journals Epitope-directed antibody selection by site-specific photocrosslinking

2020 ◽  
Vol 6 (14) ◽  
pp. eaaz7825
Author(s):  
Longxin Chen ◽  
Chaoyang Zhu ◽  
Hui Guo ◽  
Runting Li ◽  
Limeng Zhang ◽  
...  
Keyword(s):  
Phage Display ◽  
Conformational Epitope ◽  
Phage Display Library ◽  
Target Antigen ◽  
Phage Display Libraries ◽  
Antibody Phage ◽  
Antibody Phage Display ◽  
Antigen Epitope ◽  
A Site ◽  
Noncanonical Amino Acid

Currently, there are no methods available offering solutions to select and identify antibodies binding to a specific conformational epitope of an antigen. Here, we developed a method to allow epitope-directed antibody selection from a phage display library by photocrosslinking bound antibodies to a site that specifically incorporates a noncanonical amino acid, p-benzoyl-l-phenylalanine (pBpa), on the target antigen epitope. By one or two rounds of panning against antibody phage display libraries, those hits that covalently bind to the proximity site of pBpa on specific epitopes of target antigens after ultraviolet irradiation are enriched and selected. This method was applied to specific epitopes on human interleukin-1β and complement 5a. In both cases, more than one-third of hits identified bind to the target epitopes, demonstrating the feasibility and versatility of this method.

scholarly journals Antibody Cocktail Exhibits Broad Neutralization against SARS-CoV-2 and SARS-CoV-2 variants

2021 ◽  
Author(s):  
Yuanyuan Qu ◽  
Xueyan Zhang ◽  
Meiyu Wang ◽  
Lina Sun ◽  
Yongzhong Jiang ◽  
...  
Keyword(s):  
Immune Escape ◽  
Conformational Epitope ◽  
Viral Escape ◽  
Angiotensin Converting Enzyme 2 ◽  
Neutralizing Activity ◽  
High Affinity ◽  
Phage Display Libraries ◽  
Novel Variants ◽  
Antibody Phage ◽  
Antibody Phage Display

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has precipitated multiple variants resistant to therapeutic antibodies. In this study, 12 high-affinity antibodies were generated from convalescent donors in early outbreaks using immune antibody phage display libraries. Of them, two RBD-binding antibodies (F61 and H121) showed high affinity neutralization against SARS-CoV-2, whereas three S2-target antibodies failed to neutralize SARS-CoV-2. Following structure analysis, F61 identified a linear epitope located in residues G446 - S494, which overlapped with angiotensin-converting enzyme 2 (ACE2) binding sites, while H121 recognized a conformational epitope located on the side face of RBD, outside from ACE2 binding domain. Hence the cocktail of the two antibodies achieved better performance of neutralization to SARS-CoV-2. Importantly, F61 and H121 exhibited efficient neutralizing activity against variants B.1.1.7 and B.1.351, those showed immune escape. Efficient neutralization of F61 and H121 against multiple mutations within RBD revealed a broad neutralizing activity against SARS-CoV-2 variants, which mitigated the risk of viral escape. Our findings defined the basis of therapeutic cocktails of F61 and H121 with broad neutralization and delivered a guideline for the current and future vaccine design, therapeutic antibody development, and antigen diagnosis of SARS-CoV-2 and its novel variants.

Hevein-specific recombinant IgE antibodies from human single-chain antibody phage display libraries

2003 ◽  
Vol 278 (1-2) ◽  
pp. 271-281 ◽  
Author(s):  
Marja-Leena Laukkanen ◽  
Soili Mäkinen-Kiljunen ◽  
Kirsi Isoherranen ◽  
Tari Haahtela ◽  
Hans Söderlund ◽  
...  
Keyword(s):  
Phage Display ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Ige Antibodies ◽  
Phage Display Libraries ◽  
Antibody Phage ◽  
Antibody Phage Display

Single chain variable fragment antibodies against Shiga toxins isolated from a human antibody phage display library

Vaccine ◽  
2011 ◽  
Vol 29 (33) ◽  
pp. 5340-5346 ◽  
Author(s):  
Paola Neri ◽  
Naoko Shigemori ◽  
Susumu Hamada-Tsutsumi ◽  
Kentaro Tsukamoto ◽  
Hideyuki Arimitsu ◽  
...  
Keyword(s):  
Phage Display ◽  
Phage Display Library ◽  
Human Antibody ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Shiga Toxins ◽  
Antibody Phage ◽  
Antibody Phage Display

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

2020 ◽  
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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