Object's details: Characterization of mAb6-9-1 monoclonal antibody against hemagglutinin of avian influenza virus H5N1 and its engineered derivative, single-chain variable fragment antibody

Provider:Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

Description

Title:
Characterization of mAb6-9-1 monoclonal antibody against hemagglutinin of avian influenza virus H5N1 and its engineered derivative, single-chain variable fragment antibody
Creator:
Sawicka, Róża;
Siedlecki, Paweł;
Kalenik, Barbara;
Radomski, Jan P.;
Sączyńska, Violetta;
Porębska, Anna;
Szewczyk, Bogusław;
Sirko, Agnieszka;
Góra-Sochacka, Anna
Description:
Hemagglutinin (HA), as a major surface antigen of influenza virus, is widely used as a target for production of neutralizing antibodies. Monoclonal antibody, mAb6-9-1, directed against HA of highly pathogenic avian influenza virus A/swan/Poland/305-135V08/2006(H5N1) was purified from mouse hybridoma cells culture and characterized. The antigenic specificity of mAb6-9-1 was verified by testing its cross-reactivity with several variants of HA. The mimotopes recognized by mAb6-9-1 were selected from two types of phage display peptide libraries. The comparative structural model of the HA variant used for antibody generation was developed to further facilitate epitope mapping. Based on the sequences of the affinity- selected polypeptides and the structural model of HA the epitope was located to the region near the receptor binding site (RBS). Such localization of the epitope recognized by mAb6-9-1 is in concordance with its moderate hemagglutination inhibiting activity and its antigenic specificity. Additionally, total RNA isolated from the hybridoma cell line secreting mAb6-9-1 was used for obtaining two variants of cDNA encoding recombinant single-chain variable fragment (scFv) antibody. To ensure high production level and solubility in bacterial expression system, the scFv fragments were produced as chimeric proteins in fusion with thioredoxin or displayed on a phage surface after cloning into the phagemid vector. Specificity and affinity of the recombinant soluble and phage-bound scFv were assayed by suitable variants of ELISA test. The observed differences in specificity were discussed.
Object availability:
free access
Date:
2017
Type:
Article;
PeerReviewed
Language:
English;
eng
Format:
application/pdf
Relation: http://eprints.ibb.waw.pl/1375 ;http://doi.org/10.18388/abp.2016_1292 ;doi:10.18388/abp.2016_1292
Subject:
Q Science (General);
QR Microbiology;
QR180 Immunology;
QR355 Virology
Identifier: http://eprints.ibb.waw.pl/1375/1/2016_1292.pdf ;
Sawicka, Róża and Siedlecki, Paweł and Kalenik, Barbara and Radomski, Jan P. and Sączyńska, Violetta and Porębska, Anna and Szewczyk, Bogusław and Sirko, Agnieszka and Góra-Sochacka, Anna (2017) Characterization of mAb6-9-1 monoclonal antibody against hemagglutinin of avian influenza virus H5N1 and its engineered derivative, single-chain variable fragment antibody. Acta Biochimica Polonica, 64 (1). pp. 85-92. ISSN 0001-527X
Data provider:
Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

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Object's details: Characterization of mAb6-9-1 monoclonal antibody against hemagglutinin of avian influenza virus H5N1 and its engineered derivative, single-chain variable fragment antibody

Description

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