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Molecular optimization of rabies virus glycoprotein expression in Pichia pastoris.

Ben Azoun S, Belhaj AE, Göngrich R, Gasser B, Kallel H - Microb Biotechnol (2016)

Bottom Line: Nevertheless, data showed that the secretion of the optimized RABV-G gene was not tremendously increased as compared with the non-optimized one.Among these factors only PDI1, ERO1 and GPX1 proved their benefit to enhance the expression.Competitive neutralizing assay confirmed that the recombinant protein was produced in the correct conformational form in this host.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Biofermentation Unit, Institut Pasteur de Tunis, 13, place Pasteur. BP. 74, Tunis, 1002, Tunisia.

No MeSH data available.


Related in: MedlinePlus

Competition of the neutralizing activity of the rabies‐immune serum by recombinant RABV‐G. The neutralizing titres (IU ml−1) were determined in the presence of varying amounts of RABV‐G expressed by α‐7/P6 recombinant clone, NC (negative control): culture supernatant of KM71H strain transformed with empty pPICZαA vector.
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mbt212350-fig-0004: Competition of the neutralizing activity of the rabies‐immune serum by recombinant RABV‐G. The neutralizing titres (IU ml−1) were determined in the presence of varying amounts of RABV‐G expressed by α‐7/P6 recombinant clone, NC (negative control): culture supernatant of KM71H strain transformed with empty pPICZαA vector.

Mentions: To prove that the RABV‐G expressed by clone α‐7/P6 was secreted in its native form and can compete with rabies virus to react with anti‐rabies neutralizing serum and to block cell infection, the RFFIT test was used with the modifications introduced by Li et al. (2010). The neutralizing activity of rabies‐immune serum was evaluated in the presence of different levels of recombinant RABV‐G varying from 0.8 to 49 μg ml−1. As shown in Fig. 4, the neutralizing titre of the rabies‐immune serum was significantly reduced in the presence of RABV‐G compared with that seen in the absence of recombinant RABV‐G. These results suggest that the RABV‐G produced by P. pastoris was recognized by rabies virus neutralizing antibodies.


Molecular optimization of rabies virus glycoprotein expression in Pichia pastoris.

Ben Azoun S, Belhaj AE, Göngrich R, Gasser B, Kallel H - Microb Biotechnol (2016)

Competition of the neutralizing activity of the rabies‐immune serum by recombinant RABV‐G. The neutralizing titres (IU ml−1) were determined in the presence of varying amounts of RABV‐G expressed by α‐7/P6 recombinant clone, NC (negative control): culture supernatant of KM71H strain transformed with empty pPICZαA vector.
© Copyright Policy - creativeCommonsBy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835572&req=5

mbt212350-fig-0004: Competition of the neutralizing activity of the rabies‐immune serum by recombinant RABV‐G. The neutralizing titres (IU ml−1) were determined in the presence of varying amounts of RABV‐G expressed by α‐7/P6 recombinant clone, NC (negative control): culture supernatant of KM71H strain transformed with empty pPICZαA vector.
Mentions: To prove that the RABV‐G expressed by clone α‐7/P6 was secreted in its native form and can compete with rabies virus to react with anti‐rabies neutralizing serum and to block cell infection, the RFFIT test was used with the modifications introduced by Li et al. (2010). The neutralizing activity of rabies‐immune serum was evaluated in the presence of different levels of recombinant RABV‐G varying from 0.8 to 49 μg ml−1. As shown in Fig. 4, the neutralizing titre of the rabies‐immune serum was significantly reduced in the presence of RABV‐G compared with that seen in the absence of recombinant RABV‐G. These results suggest that the RABV‐G produced by P. pastoris was recognized by rabies virus neutralizing antibodies.

Bottom Line: Nevertheless, data showed that the secretion of the optimized RABV-G gene was not tremendously increased as compared with the non-optimized one.Among these factors only PDI1, ERO1 and GPX1 proved their benefit to enhance the expression.Competitive neutralizing assay confirmed that the recombinant protein was produced in the correct conformational form in this host.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Biofermentation Unit, Institut Pasteur de Tunis, 13, place Pasteur. BP. 74, Tunis, 1002, Tunisia.

No MeSH data available.


Related in: MedlinePlus