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32 results about "Factor XIII" patented technology

Factor XIII or fibrin stabilizing factor is an enzyme (EC 2.3.2.13) of the blood coagulation system that crosslinks fibrin. Deficiency of this factor (FXIIID) affects clot stability.

Process for separating proteins fibrinogen, factor XIII and biological glue from a solubilized plasma fraction and for preparing lyophilised concentrates of said proteins

ActiveUS20080207878A1Simple and rapid and low cost processEasy to optimizeFibrinogenPeptide/protein ingredientsFreeze-dryingBlood plasma
Process for separating proteins fibrinogen, Factor XIII and biological glue from a solubilized plasma fraction and for preparing lyophilised concentrates of said proteins
Summary
The invention is related to a process for separating proteins fibrinogen, Factor XIII and biological glue from a solubilized plasma fraction and for preparing freeze-dried concentrates of said proteins comprising the steps of:
    • chromatographic purification comprising the steps of loading an anion exchanger of weak base type with the said solubilized fraction, previously equilibrated with a buffer of a predetermined ionic strength of an alkaline pH, which allows to retain the biological glue, elution of the biological glue by increasing the ionic strength of the said buffer, and
    • separation of FXIII from fibrinogen by addition to at least one part of the biological glue eluate of at least one chemical agent precipitating the FXIII, and recovery of the resulting purified fibrinogen containing supernatant solution, and
    • diafiltration of the fibrinogen, biological glue and resolubilized FXIII solutions, followed by a freeze-drying of said solutions. It is also related to freeze-dried concentrates of the said proteins capable to be obtained by carrying out the process.
Owner:LABE FR DU FRACTIONNEMENT & DES BIOTECH SA

Production of in vivo N-deglycosylated recombinant proteins by co-expression with endo H

Plants have emerged as an alternative expression system and are increasingly being used byindustry and academia for producing target proteins. However, the ability of plants to glycosylate proteins can be a significant limitation for those proteins, which do not require N-glycosylation. For example, Plasmodium falciparum proteins, or A chain of human factor XIII do not carry N-linked glycans, or the protective antigen (PA) of Bacillus anthracisis not a glycoprotein; however, these proteins contain potential N-linked glycosylation sites that can be aberrantly glycosylated during expression in yeast, mammalian, or plant systems, potentially leading to reduced functionality and immunogenicity because of incorrect / altered folding and / or masking of epitopes. To overcome this problem we have recently developed a strategy of enzymatic deglycosylation of proteins in vivo by co-expressing with bacterial PNGase F (Peptide: N-glycosidase F) using transient expression in plants (WIPO Patent Application WO / 2012 / 170678), which allowed production of malaria vaccine candidate Pfs48 / 45, which can provide a high transmission blocking (TB) activity (Mamedov et al., 2012). In addition, other deglycosylated antigens induced significantly higher levels of toxin-neutralizing antibody responses in mice than compared with glycosylated forms (Mamedov et al, manuscript has been submitted). Although a PNGase F treatment (in vivodeglycosylation) removes the oligosaccharide intact, but causes amino acid change in the deglycosylated protein due to deamidation of the asparagine (N) in the NxS / T site (sequence) into an aspartate (D). In this study, a strategy was developed for production of target proteins in plants in non-N-glycosylated form, but with no amino acid change in the NxS / T site of the resulting deglycosylated proteins, which can provide production of non-N-glycosylated recombinant proteins in plants or other eukaryotic system with a native-like fold. Thus, materials and methods for in vivo de-glycosylation of recombinant N-glycosylated proteins by co-expression with Endo-β-N-acetylglucosaminidase H (Endo H) in plants, using a transient expression system are described in this invention. A method of expressing active Endo Hin plants is also provided.
Owner:MAMMEDOV TARLAN
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