scholarly journals Generation and Selection of a Panel of Pan-Filovirus Single-Chain Antibodies using Cell-Free Ribosome Display

2018 ◽  
Author(s):  
Adinarayana Kunamneni ◽  
Elizabeth C. Clarke ◽  
Chunyan Ye ◽  
Steven B. Bradfute ◽  
Ravi Durvasula
Keyword(s):  
Hemorrhagic Fever ◽  
Cross Reactivity ◽  
Free Ribosome ◽  
Rapid Diagnostics ◽  
Single Chain ◽  
Ribosome Display ◽  
Single Chain Antibodies ◽  
Single Chain Fv ◽  
New Generation ◽  
Selection Of

AbstractFiloviruses, which include ebolaviruses and marburgvirus, can cause outbreaks of highly lethal hemorrhagic fever. This disease causes significant morbidity and mortality in humans and non-human primates, with human fatality rates reaching 90% during some outbreaks. Currently, there are a lack of licensed vaccines or antivirals for these viruses. Since early symptoms of filovirus infection mimic more common diseases, there is a strong unmet public health and biodefense need for broad-spectrum filovirus rapid diagnostics. We have generated a panel of mouse single-chain Fv-antibodies (scFvs) to filovirus glycoproteins (GPs) using cell-free ribosome display and determined their cross-reactivity profiles to all known filovirus species. Two scFvs (4-2 and 22-1) were able to detect all known Ebolavirus and Marburgvirus species. This is the first report on ribosome display scFvs that can detect a broad set of filovirus GPs, which demonstrates their potential use in the development of a new generation of rapid diagnostic immunoassays.

scholarly journals Generation and Selection of a Panel of Pan-Filovirus Single-Chain Antibodies using Cell-Free Ribosome Display

2019 ◽  
Vol 101 (1) ◽  
pp. 198-206 ◽  
Author(s):  
Adinarayana Kunamneni ◽  
Elizabeth C. Clarke ◽  
Chunyan Ye ◽  
Steven B. Bradfute ◽  
Ravi Durvasula
Keyword(s):  
Free Ribosome ◽  
Single Chain ◽  
Ribosome Display ◽  
Single Chain Antibodies ◽  
Selection Of

scholarly journals Selection of Diethylstilbestrol-Specific Single-Chain Antibodies from a Non-Immunized Mouse Ribosome Display Library

PLoS ONE ◽  
2012 ◽  
Vol 7 (3) ◽  
pp. e33186 ◽  
Author(s):  
Yanan Sun ◽  
Baoan Ning ◽  
Ming Liu ◽  
Xianjun Gao ◽  
Xianjun Fan ◽  
...  
Keyword(s):  
Single Chain ◽  
Ribosome Display ◽  
Single Chain Antibodies ◽  
Selection Of

scholarly journals Correction: Selection of Diethylstilbestrol-Specific Single-Chain Antibodies from a Non-Immunized Mouse Ribosome Display Library

PLoS ONE ◽  
2012 ◽  
Vol 7 (7) ◽  
Author(s):  
Yanan Sun ◽  
Baoan Ning ◽  
Ming Liu ◽  
Xianjun Gao ◽  
Xianjun Fan ◽  
...  
Keyword(s):  
Single Chain ◽  
Ribosome Display ◽  
Single Chain Antibodies ◽  
Selection Of

Selection of bisphenol A – single-chain antibodies from a non-immunized mouse library by ribosome display

2015 ◽  
Vol 488 ◽  
pp. 59-64 ◽  
Author(s):  
Li Zhao ◽  
Baoan Ning ◽  
Jialei Bai ◽  
Xiang Chen ◽  
Yuan Peng ◽  
...  
Keyword(s):  
Bisphenol A ◽  
Single Chain ◽  
Ribosome Display ◽  
Single Chain Antibodies ◽  
Selection Of

scholarly journals Neutralizationof Enteric Coronaviruses with Escherichia coli CellsExpressing Single-Chain Fv-AutotransporterFusions

2003 ◽  
Vol 77 (24) ◽  
pp. 13396-13398 ◽  
Author(s):  
Esteban Veiga ◽  
Víctor de Lorenzo ◽  
Luis Angel Fernández
Keyword(s):  
Escherichia Coli ◽  
Infectious Agent ◽  
Target Cells ◽  
Single Chain ◽  
E Coli ◽  
Single Chain Antibodies ◽  
Single Chain Fv ◽  
Transmissible Gastroenteritis ◽  
Iga Protease

ABSTRACT We report here that fusions of single-chain antibodies (scFvs) to the autotransporter β domain of the IgA protease of Neisseria gonorrhoeae are instrumental in locating virus-neutralizing activity on the cell surface of Escherichia coli. E. coli cells displaying scFvs against the transmissible gastroenteritis coronavirus on their surface blocked in vivo the access of the infectious agent to cultured epithelial cells. This result raises prospects for antiviral strategies aimed at hindering the entry into target cells by bacteria that naturally colonize the same intestinal niches.

Selection of Single Chain Variable Fragments Specific for the Human-Inducible Costimulator Using Ribosome Display

2012 ◽  
Vol 168 (5) ◽  
pp. 967-979 ◽  
Author(s):  
Yangbin Pan ◽  
Weiping Mao ◽  
Xuanxuan Liu ◽  
Chong Xu ◽  
Zhijuan He ◽  
...  
Keyword(s):  
Single Chain ◽  
Ribosome Display ◽  
Inducible Costimulator ◽  
Single Chain Variable Fragments ◽  
Selection Of

scholarly journals Production of scFv recombinant fragments against 2,4-dichlorophenoxyacetic acid hapten using naďve phage library

2012 ◽  
Vol 48 (No. 9) ◽  
pp. 237-247 ◽  
Author(s):  
J. Brichta ◽  
H. Vesela ◽  
M. Franek
Keyword(s):  
Cross Reactivity ◽  
Phage Library ◽  
Dichlorophenoxyacetic Acid ◽  
Single Chain ◽  
Metal Affinity ◽  
Recombinant Scfv ◽  
Storage Buffer ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Recombinant Fragments ◽  
Selection Of

Three single chain variable fragment (scFv) antibodies against 2,4-dichlophenoxyacetic acid (2,4-D) herbicide were produced by the Griffin1.library. The selection of the scFv from the phage library was carried out by 2,4-D-protein coated tubes with different levels of hapten substitution in the conjugate. The scFv phage clones were isolated within the five round library panning and the antibodies were expressed in Escherichia coli HB2151. The recombinant products were purified by metal affinity chromatography yielding 200 g of pure scFv per 1 liter of bacterial culture. The antibody fragments provided steep curves in conventional indirect ELISA having the IC<sub>50</sub> values from 10.2 to 14.5 ng/ml established for 2,4-D standard. Interestingly enough, the recombinant ScFv E1 antibody exhibited 68% cross-reactivity with 2,4-dichlorphenol (2,4-D = 100%), and 38.0% with methylchlorophenoxyacetic acid (MCPA) whereas reaction with other phenoxyacetic compounds was low. Similar characteristics were obtained for other two recombinant products. Low stability for the isolated scFv antibodies was found in storage buffer even in the presence of stabilizers and protease inhibitors. Factors influencing stability of the recombinant antibodies are discussed.

scholarly journals Ribosome Display of Combinatorial Antibody Libraries Derived from Mice Immunized with Heat-Killed Xylella fastidiosa and the Selection of MopB-Specific Single-Chain Antibodies

2012 ◽  
Vol 78 (8) ◽  
pp. 2638-2647 ◽  
Author(s):  
Armaghan Azizi ◽  
Arinder Arora ◽  
Anatoliy Markiv ◽  
David J. Lampe ◽  
Thomas A. Miller ◽  
...  
Keyword(s):  
Xylella Fastidiosa ◽  
Pierce's Disease ◽  
Single Chain ◽  
Ribosome Display ◽  
Content Type ◽  
Single Chain Antibodies ◽  
Pierce’S Disease ◽  
Antibody Libraries ◽  
Potential Use

ABSTRACTPierce's disease is a devastating lethal disease ofVitus viniferagrapevines caused by the bacteriumXylella fastidiosa. There is no cure for Pierce's disease, and control is achieved predominantly by suppressing transmission of the glassy-winged sharpshooter insect vector. We present a simple robust approach for the generation of panels of recombinant single-chain antibodies against the surface-exposed elements ofX. fastidiosathat may have potential use in diagnosis and/or disease transmission blocking studies.In vitrocombinatorial antibody ribosome display libraries were assembled from immunoglobulin transcripts rescued from the spleens of mice immunized with heat-killedX. fastidiosa. The libraries were used in a single round of selection against an outer membrane protein, MopB, resulting in the isolation of a panel of recombinant antibodies. The potential use of selected anti-MopB antibodies was demonstrated by the successful application of the 4XfMopB3 antibody in an enzyme-linked immunosorbent assay (ELISA), a Western blot assay, and an immunofluorescence assay (IFA). These immortalizedin vitrorecombinant single-chain antibody libraries generated against heat-killedX. fastidiosaare a resource for the Pierce's disease research community that may be readily accessed for the isolation of antibodies against a plethora ofX. fastidiosasurface-exposed antigenic molecules.

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