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Wyświetlanie 1-3 z 3
Tytuł:
Expression, purification and characterization of glycosylated influenza H5N1 hemagglutinin produced in Pichia pastoris
Autorzy:
Kopera, Edyta
Dwornyk, Angela
Kosson, Piotr
Florys, Katarzyna
Sączyńska, Violetta
Dębski, Janusz
Cecuda-Adamczewska, Violetta
Szewczyk, Bogusław
Zagórski-Ostoja, Włodzimierz
Grzelak, Krystyna
Powiązania:
https://bibliotekanauki.pl/articles/1039268.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Avian influenza
H5N1
recombinant hemagglutinin
Pichia pastoris
Opis:
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.
Źródło:
Acta Biochimica Polonica; 2014, 61, 3; 597-602
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Expression of avian influenza haemagglutinin (H5) and chicken interleukin 2 (chIL-2) under control of the ptcB promoter in Lactococcus lactis
Autorzy:
Szatraj, Katarzyna
Szczepankowska, Agnieszka
Sączyńska, Violetta
Florys, Katarzyna
Gromadzka, Beata
Łepek, Krzysztof
Płucienniczak, Grażyna
Szewczyk, Bogusław
Zagórski-Ostoja, Włodzimierz
Bardowski, Jacek
Powiązania:
https://bibliotekanauki.pl/articles/1039270.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Lactococcus lactis
ptcB promoter
heterologous gene expression
avian influenza H5N1
H5 haemagglutinin
chicken interleukin-2
Opis:
Gram-positive and nonpathogenic lactic acid bacteria (LAB) are considered to be promising candidates for the development of new, safe systems of heterologous protein expression. Recombinant LAB has been shown to induce specific local and systemic immune response against selected pathogens, and could be a good alternative to classical attenuated carriers. The main goal of our study was to express the avian influenza haemagglutinin (H5) and chicken interleukin 2 (chIL-2) in Lactococcus lactis. Results of this study were anticipated to lead to construction of lactococcal strain(s) with potential vaccine properties against the avian influenza A (H5N1) virus. Expression of the cloned H5 gene, its His-tagged variant and chIL-2 gene, under the control of the ptcB gene promoter was attested by RT-PCR on transcriptional level and Western or dot blot analysis on translational level, demonstrating that system can be an attractive solution for production of heterologous proteins. The results of the preliminary animal trial conducted in mice are a promising step toward development of a vaccine against avian bird flu using Lactococcus lactis cells as antigen carriers.
Źródło:
Acta Biochimica Polonica; 2014, 61, 3; 609-614
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Hemagglutinin stalk domain from H5N1 strain as a potentially universal antigen
Autorzy:
Uranowska, Karolina
Tyborowska, Jolanta
Jurek, Anna
Szewczyk, Bogusław
Gromadzka, Beata
Powiązania:
https://bibliotekanauki.pl/articles/1039261.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
H5N1
universal influenza antigen
hemagglutinin stalk domain
universal influenza antibodies
FI6
Opis:
Influenza A virus infections are the major public health concern and cause significant morbidity and mortality each year worldwide. Vaccination is the main strategy of influenza epidemic prevention. However, seasonal vaccines induce strain-specific immunity and must be reformulated annually based on prediction of the strains that will circulate in the next season. Thus, it is essential to develop vaccines that would induce broad and persistent immunity to influenza viruses. Hemagglutinin is the major surface antigen of the influenza virus. Recent studies revealed the importance of HA stalk-specific antibodies in neutralization of different influenza virus strains. Therefore, it is important to design an immunogen that would focus the immune response on the HA stalk domain in order to elicit neutralizing antibodies. In the present study, we report characterization of a conserved truncated protein, potentially a universal influenza virus antigen from the H5N1 Highly Pathogenic Avian Influenza A virus strain. Our results indicate that exposure of the HA stalk domain containing conserved epitopes results in cross reactivity with different antibodies (against group 1 and 2 HAs). Additionally, we conclude that HA stalk domain contains not only conformational epitopes recognized by universal FI6 antibody, but also linear epitopes recognized by other antibodies.
Źródło:
Acta Biochimica Polonica; 2014, 61, 3; 541-550
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
    Wyświetlanie 1-3 z 3

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