scholarly journals Novel single‐chain variant of antibody against mesothelin established by phage library

2019 ◽  
Vol 110 (9) ◽  
pp. 2722-2733 ◽  
Author(s):  
Hiromasa Yakushiji ◽  
Kazuko Kobayashi ◽  
Fumiaki Takenaka ◽  
Yoshiro Kishi ◽  
Midori Shinohara ◽  
...  
Keyword(s):  
Phage Library ◽  
Single Chain ◽  
Chain Variant

scholarly journals Generation of antibody-based therapeutics targeting the idiotype of B-cell malignancies

2018 ◽  
Vol 2 (1) ◽  
pp. 12-21
Author(s):  
Emily Weiss ◽  
Robert Sarnovsky ◽  
Mitchell Ho ◽  
Evgeny Arons ◽  
Robert Kreitman ◽  
...  
Keyword(s):  
B Cell ◽  
Specific Binding ◽  
Phage Library ◽  
Selection Strategy ◽  
Proof Of Concept ◽  
Single Chain ◽  
Human Igg1 ◽  
Specificity Of Binding ◽  
Complementarity Determining Regions ◽  
Phage Selection

ABSTRACT Background A feature of many B-cell tumors is a surface-expressed immunoglobulin (sIg). The complementarity-determining regions (CDRs) of the sIg, termed the ‘idiotype’, are unique to each tumor. We report on a phage selection strategy to generate anti-idiotype therapeutics that reacts with sIg CDR3 sequences; the MEC1 B-cell tumor line was used as proof of concept. Methods To create a mimetic of the MEC1 idiotype, CDR3 sequences from heavy and light chains of the sIg were grafted into a single chain variable fragment (scFv) framework scaffold. Using the Tomlinson I phage library of human scFvs, we enriched for binders to MEC1 CDR3 sequences over unrelated CDR3 sequences. Results By ELISA we identified 10 binder phages. Of these, five were sequenced, found to be unique and characterized further. By flow cytometry each of the five phages bound to MEC1 cells, albeit with different patterns of reactivity. To establish specificity of binding and utility, the scFv sequences from two of these binders (phages 1 and 7) were converted into antibody-toxin fusion proteins (immunotoxins) and also cloned into a human IgG1 expression vector. Binders 1 and 7 immunotoxins exhibited specific killing of MEC1 cells with little toxicity for non-target B-cell lines. The full-length antibody recreated from the binder-1 scFv also exhibited specific binding. Conclusion Our results establish the utility of using engrafted CDR3 sequences for selecting phage that recognize the idiotype of B-cell tumors.

A single chain Fv derived from a filamentous phage library has distinct tumour targeting advantages over one derived from a hybridoma

1995 ◽  
Vol 61 (4) ◽  
pp. 497-501 ◽  
Author(s):  
Marlies J. Verhaar ◽  
Kerry A. Chester ◽  
Patricia A. Keep ◽  
Lynda Robson ◽  
R. Barbara Pedley ◽  
...  
Keyword(s):  
Phage Library ◽  
Filamentous Phage ◽  
Tumour Targeting ◽  
Single Chain ◽  
Single Chain Fv

scholarly journals Structural and functional characterization of an anti-West Nile virus monoclonal antibody and its single-chain variant produced in glycoengineered plants

2014 ◽  
Vol 12 (8) ◽  
pp. 1098-1107 ◽  
Author(s):  
Huafang Lai ◽  
Junyun He ◽  
Jonathan Hurtado ◽  
Jake Stahnke ◽  
Anja Fuchs ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
West Nile Virus ◽  
Functional Characterization ◽  
Single Chain ◽  
West Nile ◽  
Chain Variant

Human recombinant antibodies againstTrypanosoma cruziribosomal P2βprotein

Parasitology ◽  
2011 ◽  
Vol 138 (6) ◽  
pp. 736-747 ◽  
Author(s):  
VANINA GRIPPO ◽  
LETICIA L. NIBORSKI ◽  
KARINA A. GOMEZ ◽  
MARIANO J. LEVIN
Keyword(s):  
Lupus Erythematosus ◽  
Recombinant Antibodies ◽  
Phage Library ◽  
Single Chain ◽  
Human Mabs ◽  
Western Blots ◽  
Variable Heavy ◽  
Chagas Heart Disease ◽  
P Proteins ◽  
Ribosomal P

SUMMARYPatients with chronic Chagas' Heart Disease (cChHD) develop an antibody response that is suspected to be involved in the cardiac pathogenesis. The response againstTrypanosoma cruziribosomal P proteins is of particular interest, as these antibodies can cross-react with host cardiac receptors causing electrophysiological alterations. To better understand the humoral anti-P response we constructed a single-chain variable fragment library derived from a cChHD patient. The variable heavy and light regions were amplified from bone-marrow RNA and subcloned into the vector pComb3X. The phage library was subsequently panned againstT. cruziribosomal P2βprotein (TcP2β). We obtained 3 different human recombinant antibodies that specifically reacted with TcP2βin ELISA and Western blots. Two of them reacted with the C-terminal region of TcP2β, peptide R13, as the recombinant autoanti-P antibodies from Systemic Lupus Erythematosus (SLE) patients. Interestingly, the third one was specific for TcP2βbut did not recognize R13, confirming the specific nature of the anti-P response in Chagas disease. Neither sequence nor VH usage similarities between Chagas and SLE anti-P autoantibodies were observed. Herein, the first human mAbs against TcP2βhave been obtained and characterized showing that the humoral anti-P response is directed against the parasite and does not include an autoimmune component.

Isolation of BNYVV Coat Protein-Specific Single Chain Fv from a Mouse Phage Library Antibody

Hybridoma ◽  
2009 ◽  
Vol 28 (5) ◽  
pp. 305-313 ◽  
Author(s):  
Zahra Moghaddassi Jahromi ◽  
Ali Hatef Salmanian ◽  
Nasrin Rastgoo ◽  
Mehdi Arbabi
Keyword(s):  
Coat Protein ◽  
Phage Library ◽  
Single Chain ◽  
Single Chain Fv

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 Identification of an anti–SARS–CoV-2 receptor-binding domain–directed human monoclonal antibody from a naïve semisynthetic library

2020 ◽  
Vol 295 (36) ◽  
pp. 12814-12821 ◽  
Author(s):  
Hilal Ahmad Parray ◽  
Adarsh Kumar Chiranjivi ◽  
Shailendra Asthana ◽  
Naveen Yadav ◽  
Tripti Shrivastava ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Specific Binding ◽  
Human Monoclonal Antibody ◽  
Protein Docking ◽  
Binding Domain ◽  
Phage Library ◽  
Receptor Binding Domain ◽  
Single Chain ◽  
Phage Display Technology ◽  
Entry Points

There is a desperate need for safe and effective vaccines, therapies, and diagnostics for SARS– coronavirus 2 (CoV-2), the development of which will be aided by the discovery of potent and selective antibodies against relevant viral epitopes. Human phage display technology has revolutionized the process of identifying and optimizing antibodies, providing facile entry points for further applications. Herein, we use this technology to search for antibodies targeting the receptor-binding domain (RBD) of CoV-2. Specifically, we screened a naïve human semisynthetic phage library against RBD, leading to the identification of a high-affinity single-chain fragment variable region (scFv). The scFv was further engineered into two other antibody formats (scFv-Fc and IgG1). All three antibody formats showed high binding specificity to CoV-2 RBD and the spike antigens in different assay systems. Flow cytometry analysis demonstrated specific binding of the IgG1 format to cells expressing membrane-bound CoV-2 spike protein. Docking studies revealed that the scFv recognizes an epitope that partially overlaps with angiotensin-converting enzyme 2 (ACE2)–interacting sites on the CoV-2 RBD. Given its high specificity and affinity, we anticipate that these anti-CoV-2 antibodies will be useful as valuable reagents for accessing the antigenicity of vaccine candidates, as well as developing antibody-based therapeutics and diagnostics for CoV-2.

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