Octapharma Biopharmaceuticals GmbH

Berlin, Germany

Octapharma Biopharmaceuticals GmbH

Berlin, Germany
SEARCH FILTERS
Time filter
Source Type

Radomski K.U.,Octapharma Biopharmaceuticals GmbH | Lattner G.,R&D Plasma | Schmidt T.,Octapharma Biopharmaceuticals GmbH | Romisch J.,R&D Plasma
BioDrugs | Year: 2017

Background: The manufacturing process of a new intravenous immune globulin (IVIG) 10% liquid product incorporates two dedicated pathogen safety steps: solvent/detergent (S/D) treatment and nanofiltration (20 nm). Ion-exchange chromatography (IEC) during protein purification also contributes to pathogen safety. The ability of these three process steps to inactivate/remove viruses and prions was evaluated. Objectives: The objective of this study was to evaluate the virus and prion safety of the new IVIG 10% liquid. Methods: Bovine viral diarrhea virus (BVDV), human immunodeficiency virus type 1 (HIV-1), mouse encephalomyelitis virus (MEV), porcine parvovirus (PPV), and pseudorabies virus (PRV) were used as models for common human viruses. The hamster-adapted scrapie strain 263K (HAS 263K) was used for transmissible spongiform encephalopathies. Virus clearance capacity and robustness of virus reduction were determined for the three steps. Abnormal prion protein (PrPSc) removal and infectivity of the samples was determined. Results: S/D treatment and nanofiltration inactivated/removed enveloped viruses to below detection limits. IEC supplements viral safety and nanofiltration was highly effective in removing non-enveloped viruses and HAS 263K. Overall virus reduction factors were: ≥9.4 log10 (HIV-1), ≥13.2 log10 (PRV), ≥8.2 log10 (BVDV), ≥11.7 log10 (MEV), ≥11.6 log10 (PPV), and ≥10.4 log10 (HAS 263K). Conclusion: Two dedicated and one supplementing steps in the manufacturing process of the new IVIG 10% liquid provide a high margin of pathogen safety. © 2017 The Author(s)


Hlavica P.,Walther Straub Institute For Pharmakologie Und Toxikologie Der Lmu | Lehnerer M.,Octapharma Biopharmaceuticals GmbH
Current Drug Metabolism | Year: 2010

In view of the pivotal role played by the diversity of fatty acid-derived oxy-products in a vast array of physiological processes, precise knowledge about the molecular principles dictating substrate specificity and regioselectivity in P450-catalyzed oxidative attack on the distinctly structured carbon chains of the monocarboxylic acids is of paramount importance. Based on a general, CYP102A1-related construct, the majority of prospective key determinants participating in fatty acid recognition/binding were found to cluster near the distal heme face made up by the helical B', F, G and I tetrad as well, as the B'-C interhelical loop and certain β-sheet segments. Most of the contact sites examined show a frequency of conservation <10%, hinting at the requirement of some degree of conformational flexibility. Some decisive elements may also have a function in maintaining active-site integrity, governing substrate access to the catalytic centre, and steering the redox machinery to efficiently promote fatty acid oxidations. Physico-chemical factors imposing constraints on orientation of the fatty acid molecules towards the iron-oxene core focus on the variably expressed polarity profile of the diverse docking regions and bulkiness of critical amino acid side chains, acting as selectivity filters for the substrate homologues. Also, dynamic fluctuations of certain contact sites located in the distal backbone of P450s may impact fatty acid positioning. Genetic engineering to introduce versatile properties into fatty acid hydroxylases may give an impetus to biotechnological exploitation of the tailored enzymes in. the production of fine chemicals and therapeutic agents. © 2010 Bentham Science Publishers Ltd.


PubMed | Octapharma Biopharmaceuticals GmbH, University of Aarhus, University of Leipzig, Copenhagen University and Bioneer Biomedical Technology
Type: Journal Article | Journal: Disease models & mechanisms | Year: 2015

Animal models of familial juvenile onset of Alzheimers disease (AD) often fail to produce diverse pathological features of the disease by modification of single gene mutations that are responsible for the disease. They can hence be poor models for testing and development of novel drugs. Here, we analyze in vitro-produced stem cells and their derivatives from a large mammalian model of the disease created by overexpression of a single mutant human gene (APPsw). We produced hemizygous and homozygous radial glial-like cells following culture and differentiation of embryonic stem cells (ESCs) isolated from embryos obtained from mated hemizygous minipigs. These cells were confirmed to co-express varying neural markers, including NES, GFAP and BLBP, typical of type one radial glial cells (RGs) from the subgranular zone. These cells had altered expression of CCND1 and NOTCH1 and decreased expression of several ribosomal RNA genes. We found that these cells were able to differentiate into astrocytes upon directed differentiation. The astrocytes produced had decreased - and -secretase activity, increased -secretase activity and altered splicing of tau. This indicates novel aspects of early onset mechanisms related to cell renewal and function in familial AD astrocytes. These outcomes also highlight that radial glia could be a potentially useful population of cells for drug discovery, and that altered APP expression and altered tau phosphorylation can be detected in an in vitro model of the disease. Finally, it might be possible to use large mammal models to model familial AD by insertion of only a single mutation.


Solecka B.A.,Octapharma Biopharmaceuticals GmbH | Weise C.,Free University of Berlin | Fuchs B.,Charité - Medical University of Berlin | Kannicht C.,Octapharma Biopharmaceuticals GmbH
Thrombosis Research | Year: 2016

Introduction von Willebrand factor (VWF) is rich in cysteine; next to important structural disulfide bonds, free thiol groups are present. Free thiols on the surface of plasmatic VWF have been shown to play a role in VWF self-association and in platelet binding under pathologically high levels of shear stress. The present study explores the role of VWF free thiol groups under physiological levels of shear stress and in interactions with collagen and platelet-GPIbα receptor. Materials and methods Free and accessible thiol groups were blocked with N-ethylmaleimide (NEM) and the derivatized molecule was evaluated in functional assays. Reduced cysteine residues were identified using biotin-linked maleimide (MPB) followed by analysis of multimer and domain incorporation and by analysis of derivatized tryptic peptides by mass spectrometry. Results Blockade of free thiol groups significantly reduced VWF-mediated platelet recruitment to collagen under physiological flow conditions. This resulted from inhibition of VWF binding to both collagen and the platelet GPIb receptor. Evaluation of derivatization sites revealed a high level of derivatization in the cysteine-rich N- and C-termini of VWF. 19 MPB-derivatized peptides, 13 of which are described here for the first time, were identified by mass spectrometry. Conclusions This study shows a significant contribution of free thiol groups in VWF to the mediation of platelet adhesion under physiological shear stress conditions. The free thiol groups are shown to be involved in VWF binding to both collagen III and platelet GP1b receptor. © 2015 Elsevier Ltd. All rights reserved.


Rosenlocher J.,University of Applied Sciences, Berlin | Rosenlocher J.,Free University of Berlin | Sandig G.,University of Applied Sciences, Berlin | Kannicht C.,Octapharma Biopharmaceuticals GmbH | And 3 more authors.
Journal of Proteomics | Year: 2016

Glycosylation is the most complex post-translational modification. Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. The structural heterogeneity of glycoproteins was also described by the definition of glycoforms. We therefore introduced a new term called "glycoprotein species" to join the two concepts from different fields of biology. In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. Biological significance: Protein glycosylation is a major source for the huge amount of protein species. This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. © 2015 Elsevier B.V.


PubMed | Octapharma Biopharmaceuticals GmbH, Charité - Medical University of Berlin and University of Applied Sciences, Berlin
Type: | Journal: Journal of proteomics | Year: 2016

Glycosylation is the most complex post-translational modification. Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. The structural heterogeneity of glycoproteins was also described by the definition of glycoforms. We therefore introduced a new term called glycoprotein species to join the two concepts from different fields of biology. In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production.Protein glycosylation is a major source for the huge amount of protein species. This study extends the sight of protein species by the following contributions: 1) The new term glycoprotein species was defined to introduce the concept of glycoforms into the field. 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed.


PubMed | Octapharma Biopharmaceuticals GmbH, Free University of Berlin and Charité - Medical University of Berlin
Type: | Journal: Thrombosis research | Year: 2016

von Willebrand factor (VWF) is rich in cysteine; next to important structural disulfide bonds, free thiol groups are present. Free thiols on the surface of plasmatic VWF have been shown to play a role in VWF self-association and in platelet binding under pathologically high levels of shear stress. The present study explores the role of VWF free thiol groups under physiological levels of shear stress and in interactions with collagen and platelet-GPIb receptor.Free and accessible thiol groups were blocked with N-ethylmaleimide (NEM) and the derivatized molecule was evaluated in functional assays. Reduced cysteine residues were identified using biotin-linked maleimide (MPB) followed by analysis of multimer and domain incorporation and by analysis of derivatized tryptic peptides by mass spectrometry.Blockade of free thiol groups significantly reduced VWF-mediated platelet recruitment to collagen under physiological flow conditions. This resulted from inhibition of VWF binding to both collagen and the platelet GPIb receptor. Evaluation of derivatization sites revealed a high level of derivatization in the cysteine-rich N- and C-termini of VWF. 19 MPB-derivatized peptides, 13 of which are described here for the first time, were identified by mass spectrometry.This study shows a significant contribution of free thiol groups in VWF to the mediation of platelet adhesion under physiological shear stress conditions. The free thiol groups are shown to be involved in VWF binding to both collagen III and platelet GP1b receptor.


Dobslaff K.,University of Leipzig | Zscharnack K.,University of Leipzig | Kreisig T.,University of Leipzig | Zuchner T.,University of Leipzig | Zuchner T.,Octapharma Biopharmaceuticals GmbH
Amino Acids | Year: 2016

Immunoassays play an essential role in current research and diagnostics resulting in a variety of detection principles. Thereby, homogeneous assays are often used for a fast signal response as demanded for example in point-of-care diagnostics. These systems often rely on a competitive assay design where the sample analyte and the corresponding dye-labeled substance are competing for binding sites on an antibody present in limited amounts. Due to the similar affinities of the antibody towards the sample analyte and the competitor, both sensitivity and assay time are limited. As a consequence, a competitor with a slightly reduced affinity towards the antibody can potentially overcome these drawbacks. Here, we present the rational design of a low-affinity peptide (donor peptide) as a specific analyte competitor for a FRET-based homogeneous immunoassay for the analysis of the protein cystatin C. Thereby, the strategy of peptide-induced antibody generation was combined with the selective variation of the immunization sequence in order to achieve a lower affinity towards the antibody. We could show that shortened donor peptides improved the resulting quenching efficiency in the immunoassay. In addition, the substitution of small hydrophobic amino acids by those with a higher steric demand appeared to be the most promising strategy providing a fast assay response for cystatin C of only 90 s. © 2015 Springer-Verlag Wien.


In 2016 the World Health Organization declared the mosquito-borne Zika virus (ZIKV) a public health emergency of international concern. ZIKV is a blood-borne pathogen, which therefore causes concerns regarding the safety of human plasma-derived products due to potential contamination of the blood supply. This study investigated the effectiveness of viral inactivation steps used during the routine manufacturing of various plasma-derived products to reduce ZIKV infectivity.Human plasma and intermediates from the production of various plasma-derived products were spiked with ZIKV and subjected to virus inactivation using the identical techniques (either solvent/detergent [S/D] treatment or pasteurization) and conditions used for the actual production of the respective products. Samples were taken and the viral loads measured before and after inactivation.After S/D treatment of spiked intermediates of the plasma-derived products Octaplas(LG), Octagam, and Octanate, the viral loads were below the limit of detection in all cases. The mean log reduction factor (LRF) was at least 6.78 log for Octaplas(LG), at least 7.00 log for Octagam, and at least 6.18 log for Octanate after 60, 240, and 480 minutes of S/D treatment, respectively. For 25% human serum albumin (HSA), the mean LRF for ZIKV was at least 7.48 log after pasteurization at 60C for 120 minutes.These results demonstrate that the commonly used virus inactivation processes utilized during the production of human plasma and plasma-derived products, namely, S/D treatment or pasteurization, are effective for inactivation of ZIKV.


PubMed | Octapharma Biopharmaceuticals GmbH and University of Cologne
Type: Journal Article | Journal: Biomolecules | Year: 2016

The Drosophila melanogaster glucuronyltransferases dGlcAT-S and dGlcAT-P were reported to be expressed ubiquitously and results of in vitro activity assays indicate a functional redundancy. We analyzed both transferases in vivo and in vitro and could show significant differences in their activity towards N-and O-glycoproteins in vivo. While GlcAT-P is able to use N-linked N-acetyllactosamine chains and the O-linked T-antigen as a substrate to form non-sulfated HNK1- (GlcA1-3Gal1-4GlcNAc1-) and glucuronyl-T-antigens in vivo, GlcAT-S adds glucuronic acid only to N-linked chains, thereby synthesizing only the non-sulfated HNK1-antigen.

Loading Octapharma Biopharmaceuticals GmbH collaborators
Loading Octapharma Biopharmaceuticals GmbH collaborators