Selection of Potential Therapeutic Human Single-Chain Fv Antibodies against Cholecystokinin-B/Gastrin Receptor by Phage Display Technology

BioDrugs ◽  
2013 ◽  
Vol 27 (1) ◽  
pp. 55-67 ◽  
Author(s):  
Mohammad R. Tohidkia ◽  
Farzad Asadi ◽  
Jaleh Barar ◽  
Yadollah Omidi
Keyword(s):  
Phage Display ◽  
Single Chain ◽  
Phage Display Technology ◽  
Gastrin Receptor ◽  
Single Chain Fv ◽  
Display Technology ◽  
Selection Of

scholarly journals Selection of Linkers for a Catalytic Single-chain Antibody Using Phage Display Technology

1996 ◽  
Vol 271 (26) ◽  
pp. 15682-15686 ◽  
Author(s):  
Ying Tang ◽  
Ning Jiang ◽  
Cushrow Parakh ◽  
Donald Hilvert
Keyword(s):  
Phage Display ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Phage Display Technology ◽  
Display Technology ◽  
Selection Of

scholarly journals Characterization of LL25X, a Newly Isolated Anti-SIV Monoclonal Antibody, to Determine Binding Specificity

Elements ◽  
2017 ◽  
Vol 13 (1) ◽  
Author(s):  
Zackary Tajin Park
Keyword(s):  
Phage Display ◽  
Mammalian Cells ◽  
Expression Vector ◽  
Restriction Enzymes ◽  
Phage Display Library ◽  
Single Chain ◽  
Phage Display Technology ◽  
Mammalian Expression ◽  
Mammalian Expression Vector ◽  
Single Chain Fv

A phage display library was previously constructed from an SIV-infected rhesus macaque. Several single chain Fv (scFv), including SU24, SU343 and LL25X, were selected using phage display technology. Sequences corresponding to SU24, SU343 and LL25X were optimized for expression in a mammalian system and commercially synthesized. SU24 and SU343 had previously been cloned into a mammalian expression vector. In this study, we aimed to characterize the specificity of SU24, SU343, and LL25X.. The codon-optimized version of the scFv LL25X gene sequence was cloned into a mammalian expression vector (pCEP4).  LL25X DNA was amplified by PCR, and the PCR product and mammalian expression vector were both digested with KpnI/SapI restriction enzymes. Digested fragments were purified, and the fragments were ligated using T4DNA ligase. E. coli cells were transformed with the ligation reaction. Single colonies were selected on LB agar plates containing the selective antibiotic (ampicillin). Positive colonies were identified after DNA mini-preparation and test-digestion with KpnI and SapI. Sanger sequencing confirmed cloning results and DNA sequence accuracy. Following transfection of mammalian cells (293T), LL25X-Fc cells, and purifying our protein, the binding of LL25X-Fc to the SIV gp140 envelope protein was confirmed via ELISA and Western Blotting.

scholarly journals Characterization of Monoclonal Antibodies against Bovine herpesvirus type 1 selected by Phage Display Technology

2017 ◽  
Vol 38 (6) ◽  
pp. 3915
Author(s):  
Greice Japolla ◽  
Ana Flávia Batista Penido ◽  
Greyciele Rodrigues Almeida ◽  
Luiz Artur Mendes Bataus ◽  
Jair Pereira Cunha Junior ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Phage Display ◽  
Bovine Herpesvirus ◽  
Single Chain ◽  
Phage Display Technology ◽  
Herpesvirus Type ◽  
Wide Range ◽  
Display Technology ◽  
Bovine Herpesvirus Type 1

The specificity of monoclonal antibodies (mAbs) to desired targets makes these molecules suitable for therapeutic and diagnostic uses against a wide range of pathogens. Phage display antibody libraries offer one method by which mAbs can be selected for, without the use of conventional hybridoma technology. In this work, phage display technology was used to construct, select and characterize a combinatorial single chain fragment variable (scFv) antibody library against bovine herpesvirus type 1 (BoHV-1) from the immune repertoire of chickens immunized with the virus. In silico analysis of the hypervariable domains of the antibody heavy chains revealed a high frequency of scFv fragments with low variability, suggesting that selection had probably been carried out and favored by a few im-munogenic viral antigens. The reactivity of the scFv fragments selected against BoHV-1 was demon-strated by Phage-ELISA. A significant increase in antibody reactivity to the target was observed after six rounds of library selection, showing its potential use as a molecule for BoHV-1 diagnosis. The strategy described here opens up a field for the use of phage display as a tool for selection of mono-clonal antibodies that could be used for theranostic applications against infectious and parasitic dis-eases of veterinary interest.

A novel phage display vector for selection of target-specific peptides

2020 ◽  
Vol 33 ◽  
Author(s):  
Alex Chang ◽  
Joey P Ting ◽  
Alfonso Espada ◽  
Howard Broughton ◽  
Manuel Molina-Martin ◽  
...  
Keyword(s):  
Amino Acids ◽  
Fusion Protein ◽  
Phage Display ◽  
Copy Number ◽  
Wild Type ◽  
Phage Display Technology ◽  
Successful Isolation ◽  
Display Technology ◽  
Therapeutic Peptides ◽  
Selection Of

Abstract Intrinsic low display level of polypeptides on phage is a fundamental and limiting hurdle in successful isolation of target-specific binders by phage display technology. To circumvent this challenge, we optimized the copy number of peptides displayed on the phage surface using type 33 phage vector. We randomized the first 67 amino acids of the wild type PIII to identify mutants that would result in its reduced expression. Consequently, the display level was improved by 30-fold due to higher incorporation of the synthetic PIII–peptide fusion protein on the phage surface. Utilization of this novel phage vector should provide a solid basis for the discovery of therapeutic peptides.

scholarly journals Selection of Rabbit Single-chain Fv Fragments against the Herbicide Atrazine using a New Phage Display System

1999 ◽  
Vol 11 (1) ◽  
pp. 5-17 ◽  
Author(s):  
Yi Li ◽  
WILLIAM COCKBURN ◽  
John Kilpatrick ◽  
Garry C. Whitelam
Keyword(s):  
Phage Display ◽  
Display System ◽  
Single Chain ◽  
Single Chain Fv ◽  
Herbicide Atrazine ◽  
Selection Of

scholarly journals Generation of Novel Single-Chain Antibodies by Phage-Display Technology to Direct Imaging Agents Highly Selective to Pancreatic  - or  -Cells In Vivo

Diabetes ◽  
2009 ◽  
Vol 58 (10) ◽  
pp. 2324-2334 ◽  
Author(s):  
S. Ueberberg ◽  
J. J. Meier ◽  
C. Waengler ◽  
W. Schechinger ◽  
J. W. Dietrich ◽  
...  
Keyword(s):  
Phage Display ◽  
Imaging Agents ◽  
Single Chain ◽  
Direct Imaging ◽  
Phage Display Technology ◽  
Single Chain Antibodies ◽  
Display Technology
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