scholarly journals SINGLE CHAIN VARIABLE FRAGMENTS OF ANTIBODIES AGAINST DIPHTHERIA TOXIN B-SUBUNIT ISOLATED FROM PHAGE DISPLAY HUMAN ANTIBODY LIBRARY

2014 ◽  
Vol 7 (1) ◽  
pp. 54-59 ◽  
Author(s):  
O. S. Oliinyk ◽  
Keyword(s):  
Phage Display ◽  
Diphtheria Toxin ◽  
Human Antibody ◽  
Antibody Library ◽  
Single Chain ◽  
Toxin B ◽  
B Subunit ◽  
Single Chain Variable Fragments

Screening of Single-Chain Variable Fragments against TSP50 from a Phage Display Antibody Library and Their Expression as Soluble Proteins

2006 ◽  
Vol 11 (5) ◽  
pp. 546-552 ◽  
Author(s):  
Jingyan Wei ◽  
Yang Liu ◽  
Songchuan Yang ◽  
Junjie Xu ◽  
Hangtian Kong ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Phage Display ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sandwich Elisa ◽  
Sodium Dodecyl ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Library ◽  
Single Chain ◽  
Single Chain Variable Fragments ◽  
Phage Display Antibody

A novel gene, testes-specific protease 50 ( TSP50), is abnormally activated and differentially expressed in most patients with breast cancer, suggesting it as a novel biomarker for this disease. The possibility that TSP50 may be an oncogene is presently under investigation. In this study, the single-chain variable fragments (scFvs) against TSP50 were panned from a phage display antibody library using TSP50-specific peptide, pep-50, as a target antigen. After 4 rounds of panning, 3 clones (A1, A11, and C8) from the library were verified to show strong binding affinities for TSP50 by enzyme-linked immunosorbent assay (ELISA) and to contain the variable region genes of the light and heavy chains of scFv antibodies but different complementary determining regions by sequencing. The genes of scFv-A1 and scFv-A11 were cloned into expression vector pPELB and successfully expressed as a soluble protein in Escherichia coli Rosetta. The yields of expressions were about 4.0 to 5.0 mg of protein from 1 L of culture. The expressed proteins were purified by a 2-step procedure consisting of ion-exchange chromatography, followed by immobilized metal affinity chromatography. The purified proteins were shown a single band at the position of 31 KDa on sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Sandwich ELISA demonstrated that the expressed scFv proteins were able to specifically react with pep-50, laying a foundation for the investigation of the function of TSP50 in the development and treatment of breast cancer.

scholarly journals Obligate Multivalent Recognition of Cell Surface Tomoregulin following Selection from a Multivalent Phage Antibody Library

2006 ◽  
Vol 11 (8) ◽  
pp. 985-995 ◽  
Author(s):  
Tara Heitner ◽  
Noboru Satozawa ◽  
Kirk Mclean ◽  
David Vogel ◽  
Ronald R. Cobb ◽  
...  
Keyword(s):  
Cell Surface ◽  
Phage Display ◽  
Screening Method ◽  
Surface Protein ◽  
Binding Activity ◽  
Therapeutic Antibody ◽  
Human Antibody ◽  
Antibody Library ◽  
Single Chain ◽  
Phage Antibodies

A therapeutic antibody candidate (AT-19) isolated using multivalent phage display binds native tomoregulin (TR) as a mul-timer not as a monomer. This report raises the importance of screening and selecting phage antibodies on native antigen and reemphasizes the possibility that potentially valuable antibodies are discarded when a monomeric phage display system is used for screening. A detailed live cell panning selection and screening method to isolate multivalently active antibodies is described. AT-19 is a fully human antibody recognizing the cell surface protein TR, a proposed prostate cancer target for therapeutic antibody internalization. AT-19 was isolated from a multivalent single-chain variable fragment (scFv) antibody library rescued with hyperphage. The required multivalency for isolation of AT-19 is supported by fluorescence activated cell sorting data demonstrating binding of the multivalent AT-19 phage particles at high phage concentrations and failure of monovalent particles to bind. Pure monomeric scFv AT-19 does not bind native receptor on cells, whereas dimeric scFv or immunoglobulin G binds with nanomolar affinity. The isolation of AT-19 antibody with obligate bivalent binding activity to native TR is attributed to the use of a multivalent display of scFv on phage and the method for selecting and screening by alternate use of 2 recombinant cell lines.

scholarly journals Study on toxin-neutralization properties of recombinant single-chain variable antibody’s fragments against diphtheria toxin B subunit

2009 ◽  
Vol 25 (4) ◽  
pp. 315-318 ◽  
Author(s):  
O. S. Oliinyk ◽  
A. J. Labyntsev ◽  
N. V. Korotkevich ◽  
D. V. Kolibo ◽  
S. V. Komisarenko
Keyword(s):  
Diphtheria Toxin ◽  
Single Chain ◽  
Toxin B ◽  
B Subunit

scholarly journals Selection of antibody single-chain variable fragments with improved carbohydrate binding by phage display

1994 ◽  
Vol 269 (13) ◽  
pp. 9533-9538
Author(s):  
S.J. Deng ◽  
C.R. MacKenzie ◽  
J. Sadowska ◽  
J. Michniewicz ◽  
N.M. Young ◽  
...  
Keyword(s):  
Phage Display ◽  
Carbohydrate Binding ◽  
Single Chain ◽  
Single Chain Variable Fragments ◽  
Selection Of

scholarly journals Use of phage display for isolation and characterization of single-chain variable fragments against dihydroflavonol 4-reductase from Petunia hybrida

FEBS Letters ◽  
1997 ◽  
Vol 403 (2) ◽  
pp. 116-122 ◽  
Author(s):  
Geert De Jaeger ◽  
Emmanuel Buys ◽  
Dominique Eeckhout ◽  
Anne-Marie Bruyns ◽  
Myriam De Neve ◽  
...  
Keyword(s):  
Phage Display ◽  
Petunia Hybrida ◽  
Single Chain ◽  
Isolation And Characterization ◽  
Single Chain Variable Fragments
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