Construction and characterization of a highly reactive chicken-derived single-chain variable fragment (scFv) antibody against Staphylococcus aureus developed with the T7 phage display system

2016 ◽  
Vol 35 ◽  
pp. 149-154 ◽  
Author(s):  
Jingquan Li ◽  
Yongping Xu ◽  
Xitao Wang ◽  
Yuan Li ◽  
Lili Wang ◽  
...  
2006 ◽  
Vol 44 (2) ◽  
pp. 65 ◽  
Author(s):  
Guanxin Shen ◽  
Limin Cao ◽  
Yinchang Zhu ◽  
Weiyu Wang ◽  
Xiaorong Zhao ◽  
...  

Bioengineered ◽  
2016 ◽  
Vol 8 (5) ◽  
pp. 501-510 ◽  
Author(s):  
Hamid Nickho ◽  
Vahid Younesi ◽  
Leili Aghebati-Maleki ◽  
Morteza Motallebnezhad ◽  
Jafar Majidi Zolbanin ◽  
...  

2018 ◽  
Vol 90 (7) ◽  
pp. 1184-1191 ◽  
Author(s):  
Haixia Zhang ◽  
Xinglong Wang ◽  
Xiangrong Li ◽  
Zhongren Ma ◽  
Ruofei Feng

2021 ◽  
Author(s):  
Alexandre Figueiredo ◽  
Thiago Chaves ◽  
Fernando Conte ◽  
Rodrigo Silva ◽  
Milena Carvalho ◽  
...  

Author(s):  
Shirafkan Kordi ◽  
Mohammad Rahmati-Yamchi ◽  
Mehdi Asghari Vostakolaei ◽  
Ali Etemadie ◽  
Abolfazl Barzegari ◽  
...  

Vascular endothelial growth factor receptor 2 (VEGFR-2) is known as one of the important antigens playing a vital role in angiogenesis. In this study, phage display technology (PDT) was used to produce a single-chain variable fragment (scFv) antibody against a region of the domain 3 in VEGFR-2 called kinase insert domain receptor 3 (KDR3). After designing the KDR3 peptide and biopanning, a colony was chosen for scFv antibody expression. Following expression and purification; western blotting, dot blotting and immunofluorescence (IF) were used to evaluate the antibody function. Surface plasmon resonance (SPR) was also employed to measure affinity of produced antibody. Once a colony was selected and transferred to the expression host, the scFv antibody was expressed in the expected range of 28 kDa. Using a designed chromatography column, antibody purification was found to be about 95%. In this study, a novel scFv with the capability of binding to KDR3 was isolated and purified and its intracellular function was investigated and verified.


2012 ◽  
Vol 182 (1-2) ◽  
pp. 99-103 ◽  
Author(s):  
Dongbo Sun ◽  
Hongyan Shi ◽  
Jianfei Chen ◽  
Da Shi ◽  
Qinghe Zhu ◽  
...  

2010 ◽  
Vol 76 (24) ◽  
pp. 8184-8191 ◽  
Author(s):  
Pawan Kumar Singh ◽  
Ranu Agrawal ◽  
Dev Vrat Kamboj ◽  
Garima Gupta ◽  
M. Boopathi ◽  
...  

ABSTRACT Staphylococcal food poisoning (SFP) is one of the most prevalent causes of food-borne illness throughout the world. SFP is caused by 21 different types of staphylococcal enterotoxins produced by Staphylococcus aureus. Among these, staphylococcal enterotoxin B (SEB) is the most potent toxin and is a listed biological warfare (BW) agent. Therefore, development of immunological reagents for detection of SEB is of the utmost importance. High-affinity and specific monoclonal antibodies are being used for detection of SEB, but hybridoma clones tend to lose their antibody-secreting ability over time. This problem can be overcome by the use of recombinant antibodies produced in a bacterial system. In the present investigation, genes from a hybridoma clone encoding monoclonal antibody against SEB were immortalized using antibody phage display technology. A murine phage display library containing single-chain variable-fragment (ScFv) antibody genes was constructed in a pCANTAB 5E phagemid vector. Phage particles displaying ScFv were rescued by reinfection of helper phage followed by four rounds of biopanning for selection of SEB binding ScFv antibody fragments by using phage enzyme-linked immunosorbent assay (ELISA). Soluble SEB-ScFv antibodies were characterized from one of the clones showing high affinity for SEB. The anti-SEB ScFv antibody was highly specific, and its affinity constant was 3.16 nM as determined by surface plasmon resonance (SPR). These results demonstrate that the recombinant antibody constructed by immortalizing the antibody genes from a hybridoma clone is useful for immunodetection of SEB.


Author(s):  
Xiangying Deng ◽  
Li Wang ◽  
Xiaolong You ◽  
Pei Dai ◽  
Yanhua Zeng

Sign in / Sign up

Export Citation Format

Share Document