scholarly journals Selection and characterization of scFv antibody against nucleocapsid protein of Porcine reproductive and respiratory syndrome virus

2020 ◽  
Vol 89 (1) ◽  
pp. 39-45
Author(s):  
Magdalena Krasna ◽  
Vladimír Celer
Keyword(s):  
Infectious Agent ◽  
Scfv Antibody ◽  
Respiratory Syndrome Virus ◽  
Single Chain ◽  
Single Chain Antibody ◽  
N Protein ◽  
E Coli ◽  
Metal Affinity ◽  
Binding Epitope

Porcine reproductive and respiratory syndrome virus (PRRSV) is a widespread infectious agent in pigs. Nucleocapsid (N) protein of PRRSV has been identified as the most immunodominant viral protein. The main goal of the work was the selection and characterization of a single-chain antibody fragments (scFv) antibody specific to the N protein. Specific scFv antibody clone D5 was selected from the Tomlinson phagemid library and purified by immobilized metal affinity chromatography from the periplasmatic space of E. coli cells. The antibody was then characterized by sequencing and the ability to recognize the native virus N protein by Western blot and competitive ELISA. Pepscan analysis identified the position of the binding epitope between amino acids 62–84 of the N protein. Our study could help to improve the diagnostics and prevention of PRRSV in Central Europe.

scholarly journals Recombinant scFv Antibodies against E Protein and N Protein of Severe Acute Respiratory Syndrome Virus

2004 ◽  
Vol 36 (8) ◽  
pp. 541-547 ◽  
Author(s):  
Hui Liu ◽  
Yan-Li Ding ◽  
Wei Han ◽  
Mei-Yun Liu ◽  
Rui-Yang Tian ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Binding Activity ◽  
Scfv Antibody ◽  
Respiratory Syndrome Virus ◽  
Single Chain ◽  
N Protein ◽  
Recombinant Scfv ◽  
E Protein ◽  
Equilibrium Dissociation ◽  
Site Binding

Abstract Three single chain antibodies (scFv) against the proteins of severe acute respiratory syndrome coronavirus (SARS-CoV) were isolated by phage display from an scFv antibody library. Bio-panning was carried out against immobilized purified envelope (E) and nucleocapsid (N) proteins of SARS-CoV. Their binding activity and specificity to E or N protein of SARS-CoV were characterized by phage-ELISA. Two of them, B10 and C20, could recognize non-overlapping epitopes of the E protein according to the two-site binding test result. Clone A17 could recognize N protein. The sequence of the epitope or overlapping epitope of scFv antibody A17 was PTDSTDNNQNGGRNGARPKQRRPQ. The affinity (equilibrium dissociation constant, Kd) of SARS-CoV E protein was 5.7×10−8 M for B10 and 8.9×10−8 M for C20. The affinity of A17 for N protein was 2.1×10−6 M. All three scFv antibodies were purified with affinity chromatography and determined by Western blot.

scholarly journals Preparation and Characterization of Paclitaxel-loaded PLGA Nanoparticles Coated with Cationic SM5-1 Single-chain Antibody

BMB Reports ◽  
2007 ◽  
Vol 40 (5) ◽  
pp. 731-739 ◽  
Author(s):  
Geng Kou ◽  
Jie Gao ◽  
Hao Wang ◽  
Huaiwen Chen ◽  
Bohua Li ◽  
...  
Keyword(s):  
Plga Nanoparticles ◽  
Single Chain ◽  
Single Chain Antibody

Isolation by phage display and characterization of a single-chain antibody specific for O6-methyldeoxyguanosine

2001 ◽  
Vol 46 (12) ◽  
pp. 1024-1029 ◽  
Author(s):  
Heping Dai ◽  
Rouhong Zhao ◽  
Haug Brenda ◽  
Sean M. Hemmingsen ◽  
Wei Xiao
Keyword(s):  
Phage Display ◽  
Single Chain ◽  
Single Chain Antibody

Characterization of a unique human single-chain antibody isolated by phage-display selection on membrane-bound mosquito midgut antigens

2002 ◽  
Vol 261 (1-2) ◽  
pp. 73-83 ◽  
Author(s):  
Brian D Foy ◽  
Gerry F Killeen ◽  
Ross H Frohn ◽  
Daniel Impoinvil ◽  
Andrew Williams ◽  
...  
Keyword(s):  
Phage Display ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Mosquito Midgut ◽  
Midgut Antigens ◽  
Membrane Bound

scholarly journals Characterization of the first-in-class T-cell-engaging bispecific single-chain antibody for targeted immunotherapy of solid tumors expressing the oncofetal protein claudin 6

2015 ◽  
Vol 5 (3) ◽  
pp. e1091555 ◽  
Author(s):  
Christiane R. Stadler ◽  
Hayat Bähr-Mahmud ◽  
Laura M. Plum ◽  
Kathrin Schmoldt ◽  
Anne C. Kölsch ◽  
...  
Keyword(s):  
T Cell ◽  
Solid Tumors ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Targeted Immunotherapy
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