Object's details: Expression, purification and characterization of glycosylated influenza H5N1 hemagglutinin produced in Pichia pastoris.

Provider:Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

Description

Title:
Expression, purification and characterization of glycosylated influenza H5N1 hemagglutinin produced in Pichia pastoris.
Creator:
Kopera, Edyta;
Dvornyk, Angela;
Kosson, Piotr;
Florys, Katarzyna;
Sączyńska, Violetta;
Dębski, Janusz;
Cecuda-Adamczewska, Violetta;
Szewczyk, Bogusław;
Zagórski-Ostoja, Włodzimierz;
Grzelak, Krystyna
Description:
The A/swan/Poland/305-135V08/2006 (H5N1-subtype) hemagglutinin (HA) gene was cloned and expressed in yeast Pichia pastoris (P. pastoris). The HA cDNA lacking the C-terminal transmembrane anchor-coding sequence was fused to an α-factor leader peptide and placed under control of the methanol-inducible P. pastoris alcohol oxidase 1 (AOX1) promoter. Two P. pastoris strains: SMD 1168 and KM 71 were used for protein expression. Recombinant HA protein was secreted into the culture medium reaching an approximately 15 mg/L (KM 71 strain). Fusion protein with a His6 tag was purified to homogeneity in one step affinity chromatography. SDS-PAGE and MS/MS analysis indicated that the protein is cleaved into HA1 and HA2 domains linked by a disulfide bond. Analysis of the N-linked glycans revealed that the overexpressed HA is fully glycosylated at the same sites as the native HA in the vaccine strain. Immunological activity of the hemagglutinin protein was tested in mice, where rHA elicited a high immune response.
Object availability:
free access
Date:
2014
Type:
Article;
PeerReviewed
Language:
English
Format:
application/pdf
Relation: http://eprints.ibb.waw.pl/799 ;http://www.actabp.pl/pdf/3_2014/597.pdf
Subject:
QR180 Immunology;
QR355 Virology
Identifier: http://eprints.ibb.waw.pl/799/1/EK_ABP.pdf ;
Kopera, Edyta and Dvornyk, Angela and Kosson, Piotr and Florys, Katarzyna and Sączyńska, Violetta and Dębski, Janusz and Cecuda-Adamczewska, Violetta and Szewczyk, Bogusław and Zagórski-Ostoja, Włodzimierz and Grzelak, Krystyna (2014) Expression, purification and characterization of glycosylated influenza H5N1 hemagglutinin produced in Pichia pastoris. Acta biochimica Polonica, 61 (3). pp. 597-602. ISSN 1734-154X
Data provider:
Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

Similar objects

Expression, purification and characterization of glycosylated influenza H5N1 hemagglutinin produced in Pichia pastoris.

Date:2014

Type:other

Immunogenicity of DNA Vaccine against H5N1 Containing Extended Kappa B Site: In Vivo Study in Mice and Chickens

Date:2017.08.24

Type:other

A prime/boost vaccination with HA DNA and Pichia -produced HA protein elicits a strong humoral response in chickens against H5N1

Date:2017.01.31

Type:other

Virus-like particles as vaccine.

Creator:Chroboczek, Jadwiga | Szurgot, Inga | Szolajska, Ewa

Date:2014

Type:other

Polish Vaccine Consortium--a new national player in the influenza research.

Date:2014

Type:other

Interactions of Bacteriophages with Animal and Human Organisms—Safety Issues in the Light of Phage Therapy

Date:2021.08.19

Type:other

Phage Therapy vs. the Use of Antibiotics in the Treatment of Salmonella-Infected Chickens: Comparison of Effects on Hematological Parameters and Selected Biochemical Markers

Creator:Grabowski, Łukasz | Węgrzyn, Grzegorz | Węgrzyn, Alicja | Podlacha, Magdalena

Date:2022.12.09

Type:other

Highly different effects of phage therapy and antibiotic therapy on immunological responses of chickens infected with Salmonella enterica serovar Typhimurium

Creator:Grabowski, Łukasz | Węgrzyn, Grzegorz | Węgrzyn, Alicja | Podlacha, Magdalena

Date:2022.09.23

Type:other

Federacja Bibliotek Cyfrowych

Object's details: Expression, purification and characterization of glycosylated influenza H5N1 hemagglutinin produced in Pichia pastoris.

Description

Similar objects