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

scholarly journals Generation and Characterization of a Monoclonal Antibody Against prM Protein of West Nile Virus

2014 ◽  
Vol 33 (6) ◽  
pp. 438-443 ◽  
Author(s):  
Li-Ping Guo ◽  
Hong Huo ◽  
Xiao-Lei Wang ◽  
Zhi-Gao Bu ◽  
Rong-Hong Hua
Keyword(s):  
Monoclonal Antibody ◽  
West Nile Virus ◽  
West Nile ◽  
Prm Protein

scholarly journals Structural and functional characterization of the single-chain Fv fragment from a unique HCV E1E2-specific monoclonal antibody

FEBS Letters ◽  
2013 ◽  
Vol 587 (20) ◽  
pp. 3335-3340
Author(s):  
Catherine Fallecker ◽  
Nicolas Tarbouriech ◽  
Mohammed Habib ◽  
Marie-Anne Petit ◽  
Emmanuel Drouet
Keyword(s):  
Monoclonal Antibody ◽  
Functional Characterization ◽  
Specific Monoclonal Antibody ◽  
Single Chain ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment

scholarly journals Generation and functional characterization of a single-chain variable fragment (scFv) of the anti-FGF2 3F12E7 monoclonal antibody

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Rodrigo Barbosa de Aguiar ◽  
Tábata de Almeida da Silva ◽  
Bruno Andrade Costa ◽  
Marcelo Ferreira Marcondes Machado ◽  
Renata Yoshiko Yamada ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Functional Characterization ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Variable Regions ◽  
Recombinant Scfv ◽  
Antitumor Potential ◽  
Single Chain Variable Fragments ◽  
Binding Determinants

AbstractSingle-chain variable fragments (scFvs) are small-sized artificial constructs composed of the immunoglobulin heavy and light chain variable regions connected by a peptide linker. We have previously described an anti-fibroblast growth factor 2 (FGF2) immunoglobulin G (IgG) monoclonal antibody (mAb), named 3F12E7, with notable antitumor potential revealed by preclinical assays. FGF2 is a known angiogenesis-associated molecule implicated in tumor progression. In this report, we describe a recombinant scFv format for the 3F12E7 mAb. The results demonstrate that the generated 3F12E7 scFv, although prone to aggregation, comprises an active anti-FGF2 product that contains monomers and small oligomers. Functionally, the 3F12E7 scFv preparations specifically recognize FGF2 and inhibit tumor growth similar to the corresponding full-length IgG counterpart in an experimental model. In silico molecular analysis provided insights into the aggregation propensity and the antigen-recognition by scFv units. Antigen-binding determinants were predicted outside the most aggregation-prone hotspots. Overall, our experimental and prediction dataset describes an scFv scaffold for the 3F12E7 mAb and also provides insights to further engineer non-aggregated anti-FGF2 scFv-based tools for therapeutic and research purposes.

Functional Characterization of PM6/13, a β3-specific (GPIIIa/CD61) Monoclonal Antibody that Shows Preferential Inhibition of Fibrinogen Binding over Fibronectin Binding to Activated Human Platelets

1998 ◽  
Vol 79 (01) ◽  
pp. 177-185 ◽  
Author(s):  
Ashia Siddiqua ◽  
Michael Wilkinson ◽  
Vijay Kakkar ◽  
Yatin Patel ◽  
Salman Rahman ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Platelet Aggregation ◽  
Platelet Rich Plasma ◽  
Functional Characterization ◽  
Human Platelets ◽  
Western Blots ◽  
Activated Platelets ◽  
Reducing Conditions ◽  
Fibronectin Binding

SummaryWe report the characterization of a monoclonal antibody (MAb) PM6/13 which recognises glycoprotein IIIa (GPIIIa) on platelet membranes and in functional studies inhibits platelet aggregation induced by all agonists examined. In platelet-rich plasma, inhibition of aggregation induced by ADP or low concentrations of collagen was accompanied by inhibition of 5-hydroxytryptamine secretion. EC50 values were 10 and 9 [H9262]g/ml antibody against ADP and collagen induced responses respectively. In washed platelets treated with the cyclooxygenase inhibitor, indomethacin, PM6/13 inhibited platelet aggregation induced by thrombin (0.2 U/ml), collagen (10 [H9262]g/ml) and U46619 (3 [H9262]M) with EC50 = 4, 8 and 4 [H9262]g/ml respectively, without affecting [14C]5-hydroxytryptamine secretion or [3H]arachidonate release in appropriately labelled cells. Studies in Fura 2-labelled platelets revealed that elevation of intracellular calcium by ADP, thrombin or U46619 was unaffected by PM6/13 suggesting that the epitope recognised by the antibody did not influence Ca2+ regulation. In agreement with the results from the platelet aggregation studies, PM6/13 was found to potently inhibit binding of 125I-fibrinogen to ADP activated platelets. Binding of this ligand was also inhibited by two other MAbs tested, namely SZ-21 (also to GPIIIa) and PM6/248 (to the GPIIb-IIIa complex). However when tested against binding of 125I-fibronectin to thrombin stimulated platelets, PM6/13 was ineffective in contrast with SZ-21 and PM6/248, that were both potent inhibitors. This suggested that the epitopes recognised by PM6/13 and SZ-21 on GPIIIa were distinct. Studies employing proteolytic dissection of 125I-labelled GPIIIa by trypsin followed by immunoprecipitation with PM6/13 and analysis by SDS-PAGE, revealed the presence of four fragments at 70, 55, 30 and 28 kDa. PM6/13 did not recognize any protein bands on Western blots performed under reducing conditions. However Western blotting analysis with PM6/13 under non-reducing conditions revealed strong detection of the parent GP IIIa molecule, of trypsin treated samples revealed recognition of an 80 kDa fragment at 1 min, faint recognition of a 60 kDa fragment at 60 min and no recognition of any product at 18 h treatment. Under similar conditions, SZ-21 recognized fragments at 80, 75 and 55 kDa with the 55kDa species persisting even after 18 h trypsin treatment. These studies confirm the epitopes recognised by PM6/13 and SZ-21 to be distinct and that PM6/13 represents a useful tool to differentiate the characteristics of fibrinogen and fibronectin binding to the GPIIb-IIIa complex on activated platelets.

Sign in / Sign up

Export Citation Format

Share Document