Crucell Terug naar discussie overzicht

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1. flosz 24 september 2008 20:18

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   First administration to humans of a monoclonal antibody cocktail against rabies virus: Safety, tolerability, and neutralizing activity.
   
   2008 Sep 16
   Crucell Holland BV, Leiden, The Netherlands.
   
   Immediate passive immune prophylaxis as part of rabies post-exposure prophylaxis (PEP) often cannot be provided due to limited availability of human or equine rabies immunoglobulin (HRIG and ERIG, respectively). We report first clinical data from two phase I studies evaluating a monoclonal antibody cocktail CL184 against rabies. The studies included healthy adult subjects in the USA and India and involved two parts. First, subjects received a single intramuscular dose of CL184 or placebo in a double blind, randomized, dose-escalation trial. Second, open-label CL184 (20IU/kg) was co-administered with rabies vaccine. Safety was the primary objective and rabies virus neutralizing activity (RVNA) was investigated as efficacy parameter. Pain at the CL184 injection site was reported by less than 40% of subjects; no fever or local induration, redness or swelling was observed. RVNA was detectable from day 1 to day 21 after a single dose of CL184 20 or 40IU/kg. All subjects had adequate (>0.5IU/mL) RVNA levels from day 14 onwards when combined with rabies vaccine. CL184 appears promising as an alternative to RIG in PEP.
   
   www.ncbi.nlm.nih.gov/pubmed/18804136?...
   

   “Keep on Rockin” go!
   www.youtube.com/watch?v=OF1kCpCl9a4
   
2. flosz 24 oktober 2008 19:06

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   J Virol. 2008 Oct 22.
   
   The effect of neutralising sera on factor X-mediated adenoviral serotype 5 gene transfer.
   
   Parker AL, Waddington SN, Buckley SM, Custers J, Havenga MJ, van Rooijen N, Goudsmit J, McVey JH, Nicklin SA, Baker AH.
   British Heart Foundation Glasgow Cardiovascular Research Centre, University of Glasgow, 126 University Place, Glasgow, G12 8TA, UK; Department of Haematology, Haemophilia Centre and Thrombosis Unit, Royal Free and University College Medical School, Rowland Hill Street, London, NW3 2PF, UK; Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands; TNO Biosciences, PO Box 2215, 2301CE, Leiden, The Netherlands; Department of Molecular Cell Biology, Vrije Universiteit Medical Center (VUMC), 1081 BT Amsterdam, The Netherlands; Thrombosis Research Institute, Manresa Road, London, SW3 6LR, UK.
   
   The deployment of Adenovirus serotype 5 (Ad5) based vectors is hampered by pre-existing immunity. When delivered intravenously, hepatocyte transduction is mediated by the hexon:coagulation factor (F)X interaction. Here, we demonstrate that human sera efficiently block FX-mediated cellular binding and transduction of Ad5 based vectors in vitro. Neutralising activity correlated well with the ability to inhibit Ad5 mediated liver transduction, suggesting pre-screening patient sera in this manner accurately predicts the efficacy of Ad5 based gene therapies. Neutralisation in vitro can be partially bypassed by pseudotyping with Ad45 fiber protein, indicating a proportion of neutralising antibodies are directed against the Ad5 fiber.
   PMID: 18945780
   
   www.ncbi.nlm.nih.gov/pubmed/18945780?...
   
3. flosz 24 oktober 2008 19:13

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   Zie ook(uit feb., 2008):
   
   Cell. 2008 Feb 8;132(3):397-409.
   Adenovirus serotype 5 hexon mediates liver gene transfer.
   Waddington SN, McVey JH, Bhella D, Parker AL, Barker K, Atoda H, Pink R, Buckley SM, Greig JA, Denby L, Custers J, Morita T, Francischetti IM, Monteiro RQ, Barouch DH, van Rooijen N, Napoli C, Havenga MJ, Nicklin SA, Baker AH.
   Department of Haematology, Haemophilia Centre and Haemostasis Unit, Royal Free and University College Medical School, London NW3 2PF, UK.
   
   Adenoviruses are used extensively as gene transfer agents, both experimentally and clinically. However, targeting of liver cells by adenoviruses compromises their potential efficacy. In cell culture, the adenovirus serotype 5 fiber protein engages the coxsackievirus and adenovirus receptor (CAR) to bind cells. Paradoxically, following intravascular delivery, CAR is not used for liver transduction, implicating alternate pathways. Recently, we demonstrated that coagulation factor (F)X directly binds adenovirus leading to liver infection. Here, we show that FX binds to the Ad5 hexon, not fiber, via an interaction between the FX Gla domain and hypervariable regions of the hexon surface. Binding occurs in multiple human adenovirus serotypes. Liver infection by the FX-Ad5 complex is mediated through a heparin-binding exosite in the FX serine protease domain. This study reveals an unanticipated function for hexon in mediating liver gene transfer in vivo.
   PMID: 18267072
   www.ncbi.nlm.nih.gov/pubmed/18267072?...
   
4. flosz 24 oktober 2008 19:24

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   Nature. 2008 Oct 2;455(7213):613-9. Links
   Challenges in the development of an HIV-1 vaccine.
   
   Barouch DH.
   Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA. dbarouch@bidmc.harvard.edu
   
   The development of a safe and effective human immunodeficiency virus (HIV)-1 vaccine is a critically important global health priority. Despite recent advances in our understanding of HIV-1 pathogenesis and immunology, however, major scientific obstacles remain. Prototype HIV-1 vaccine candidates aimed at eliciting humoral and cellular immune responses have so far failed to protect against HIV-1 infection or to reduce viral loads after infection in clinical efficacy studies. A renewed and coordinated commitment to basic discovery research, preclinical studies and clinical trials will therefore be required to overcome the hurdles currently facing the field. Here I review key challenges and future prospects in the quest to develop a prophylactic HIV-1 vaccine.
   PMID: 18833271
   www.ncbi.nlm.nih.gov/pubmed/18833271?
   ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum
   
   www.nature.com/nature/journal/v455/n7...
   *************************
   Jun., 2008:
   Nature Medicine 14, 617-621 (1 June 2008) | doi:10.1038/nm.f.1759;
   Nonhuman primate models and the failure of the Merck HIV-1 vaccine in humans
   David I Watkins , Dennis R Burton , Esper G Kallas , John P Moore & Wayne C Koff
   Abstract
   The adenovirus type 5 (Ad5)-based vaccine developed by Merck failed to either prevent HIV-1 infection or suppress viral load in subsequently infected subjects in the STEP human Phase 2b efficacy trial. Analogous vaccines had previously also failed in the simian immunodeficiency virus (SIV) challenge–rhesus macaque model. In contrast, vaccine protection studies that used challenge with a chimeric simian-human immunodeficiency virus (SHIV89.6P) in macaques did not predict the human trial results. Ad5 vector–based vaccines did not protect macaques from infection after SHIV89.6P challenge but did cause a substantial reduction in viral load and a preservation of CD4+ T cell counts after infection, findings that were not reproduced in the human trials. Although the SIV challenge model is incompletely validated, we propose that its expanded use can help facilitate the prioritization of candidate HIV-1 vaccines, ensuring that resources are focused on the most promising candidates. Vaccine designers must now develop T cell vaccine strategies that reduce viral load after heterologous challenge.
   www.nature.com/nm/journal/v14/n6/full...
   
5. [verwijderd] 21 november 2008 16:42

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   Published online 2008 November 21. doi: 10.1371/journal.pone.0003790.
   
   rBCG Induces Strong Antigen-Specific T Cell Responses in Rhesus Macaques in a Prime-Boost Setting with an Adenovirus 35 Tuberculosis Vaccine Vector
   Isabelle Magalhaes,1,2 Donata R. Sizemore,3 Raija K. Ahmed,2 Stefanie Mueller,3 Lena Wehlin,2 Charles Scanga,3 Frank Weichold,3 Giulia Schirru,4 Maria Grazia Pau,4 Jaap Goudsmit,4 Sharon Kühlmann-Berenzon,2 Mats Spångberg,2 Jan Andersson,5 Hans Gaines,2 Rigmor Thorstensson,2 Yasir A. W. Skeiky,3 Jerry Sadoff,3 and Markus Maeurer1,2*
   1Microbiology, Tumor and Cell Biology Center, Karolinska Institutet, Solna, Sweden
   2The Swedish Institute for Infectious Disease Control, Solna, Sweden
   3Aeras Global TB Vaccine Foundation, Rockville, Maryland, United States of America
   4Crucell Holland BV, Leiden, The Netherlands
   5Department of Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, Stockholm, Sweden
   Jacques Zimmer, Editor
   Centre de Recherche Public-Santé, Luxembourg
   www.pubmedcentral.nih.gov/articlerend...
6. [verwijderd] 21 november 2008 19:00

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   Nature. Over HIV vaccinatie met Dan Barouch's groep:
   (voor de volledigheid)
   
   www.nature.com/nature/journal/vaop/nc...
   
   Immune control of an SIV challenge by a T-cell-based vaccine in rhesus monkeys
   
   Jinyan Liu1, Kara L. O'Brien1, Diana M. Lynch1, Nathaniel L. Simmons1, Annalena La Porte1, Ambryice M. Riggs1, Peter Abbink1, Rory T. Coffey1, Lauren E. Grandpre1, Michael S. Seaman1, Gary Landucci2, Donald N. Forthal2, David C. Montefiori3, Angela Carville4, Keith G. Mansfield4, Menzo J. Havenga5, Maria G. Pau6, Jaap Goudsmit6 & Dan H. Barouch1
   
   1. Division of Viral Pathogenesis, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA
   2. University of California, Irvine School of Medicine, Irvine, California 92697, USA
   3. Duke University Medical Center, Durham, North Carolina 27710, USA
   4. New England Primate Research Center, Southborough, Massachusetts 01772, USA
   5. TNO Biosciences, 2301 CE, Leiden, The Netherlands
   6. Crucell Holland BV, 2301 CA, Leiden, The Netherlands
   
   A recombinant adenovirus serotype 5 (rAd5) vector-based vaccine for HIV-1 has recently failed in a phase 2b efficacy study in humans1, 2. Consistent with these results, preclinical studies have demonstrated that rAd5 vectors expressing simian immunodeficiency virus (SIV) Gag failed to reduce peak or setpoint viral loads after SIV challenge of rhesus monkeys (Macaca mulatta) that lacked the protective MHC class I allele Mamu-A*01 (ref. 3). Here we show that an improved T-cell-based vaccine regimen using two serologically distinct adenovirus vectors afforded substantially improved protective efficacy in this challenge model. In particular, a heterologous rAd26 prime/rAd5 boost vaccine regimen expressing SIV Gag elicited cellular immune responses with augmented magnitude, breadth and polyfunctionality as compared with the homologous rAd5 regimen. After SIVMAC251 challenge, monkeys vaccinated with the rAd26/rAd5 regimen showed a 1.4 log reduction of peak and a 2.4 log reduction of setpoint viral loads as well as decreased AIDS-related mortality as compared with control animals. These data demonstrate that durable partial immune control of a pathogenic SIV challenge for more than 500 days can be achieved by a T-cell-based vaccine in Mamu-A*01-negative rhesus monkeys in the absence of a homologous Env antigen. These findings have important implications for the development of next-generation T-cell-based vaccine candidates for HIV-1
7. flosz 30 december 2008 20:24

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   Van Solvay.
   Vaccine. 2008 Dec 22.
   HPLC-based quantification of haemagglutinin in the production of egg- and MDCK cell-derived influenza virus seasonal and pandemic vaccines.
   
   Kapteyn JC, Porre AM, de Rond EJ, Hessels WB, Tijms MA, Kessen H, Slotboom AM, Oerlemans MA, Smit D, van der Linden J, Schoen P, Thus JL.
   Solvay Biologicals, C.J. van Houtenlaan 36, 1381 CP, Weesp, The Netherlands.
   
   The haemagglutinin (HA) content is an important specification of influenza vaccines. Recently, a reversed-phase high performance liquid chromatography (RP-HPLC) method for quantification of HA in PER.C6((R)) cell culture-based whole virus vaccines has been reported, having a high sensitivity, precision, broad range, and high sample throughput [Kapteyn JC, Drissi Saidi M, Dijkstra R, Kars C, Tjon CMS-K, Weverling GJ et al. Haemagglutinin quantification and identification of influenza A&B strains propagated in PER.C6((R)) cells: a novel RP-HPLC method. Vaccine 2006;24:3137-44]. This RP-HPLC assay is based on measuring the peak area of HA1, the hydrophilic subunit of HA, which turned out to be proportional to the amount of HA analyzed. Here, we present data demonstrating that this RP-HPLC method is also highly suitable for HA quantification of active and BPL- or formaldehyde-inactivated egg-based and MDCK cell-based whole virus samples, including egg allantoic harvest, and in final (monovalent) subunit vaccines, including those for pandemic H5N1 strains and for virosomal vaccines. In addition, the RP-HPLC assay was demonstrated to be a very powerful tool in the early stages of seasonal influenza vaccine production, when homologous serial radial immunodiffusion (SRID) reagents are not yet available, enabling fast and reliable viral growth studies in eggs in order to select the best growing virus strains or reassortants for the production of the seasonal trivalent influenza vaccine. Because of its high sensitivity, the RP-HPLC assay has shown its enormous value in supporting small scale MDCK-based (H5N1) influenza virus production models. Finally, the observed differences between HA1 molecules from various HA subtypes in UV absorbance, FLD response, and in the actual retention times in RP-HPLC are discussed in relation to the primary structure of the HA1 molecules studied.
   PMID: 19110022
   www.ncbi.nlm.nih.gov
8. flosz 2 januari 2009 11:02

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   Vaccine. 2008 Dec 26.
   Safe vaccination of children with a virosomal adjuvanted influenza vaccine.
   
   Zie post MeawandMoo1 - 1 jan 09, 21:38
   www.iex.nl/forum/topic.asp?forum=228&...
   
9. flosz 18 januari 2009 12:48

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   Vaccine. 2009 Jan 12
   Enhanced immunogenicity of an oral inactivated cholera vaccine in infants in Bangladesh obtained by zinc supplementation and by temporary withholding breast-feeding.
   
   Ahmed T, Svennerholm AM, Tarique AA, Sultana GN, Qadri F.
   International Centre for Diarrhoeal Disease Research, Bangladesh, GPO Box 128, Dhaka 1000, Bangladesh; Gothenburg University Vaccine Research Institute (GUVAX) and Department of Microbiology and Immunology, The Sahlgrenska Academy at University of Gothenburg, Box 435 S-40530, Göteborg, Sweden.
   
   The killed oral cholera vaccine Dukoral is recommended for adults and only children over 2 years of age, although cholera is seen frequently in younger children and there is an urgent need for a vaccine for them. Since decreased immunogenicity of oral vaccines in children in developing countries is a critical problem, we tested interventions to enhance responses to Dukoral. We evaluated the effect on the immune responses by temporarily withholding breast-feeding or by giving zinc supplementation. Two doses of Dukoral consisting of killed cholera vibrios and cholera B subunit were given to 6-18 months old Bangladeshi children (n=340) and safety and immunogenicity studied. Our results showed that two doses of the vaccine were safe and induced antibacterial (vibriocidal) antibody responses in 57% and antitoxin responses in 85% of the children. Immune responses were comparable after intake of one and two doses. Temporary withholding breast-feeding for 3h before immunization or supplementation with 20mg of zinc per day for 42 days resulted in increased magnitude of vibriocidal antibodies (77% and 79% responders, respectively). Administration of vaccines without buffer or in water did not result in reduction of vibriocidal responses. This study demonstrates that the vaccine is safe and immunogenic in children under 2 years of age and that simple interventions can enhance immune responses in young children.
   PMID: 19146904
   
   www.ncbi.nlm.nih.gov/pubmed/19146904?
   
   Vaccine. 2009 Jan 12.
   
   Robust gut associated vaccine-specific antibody-secreting cell responses are detected at the mucosal surface of Bangladeshi subjects after immunization with an oral killed bivalent V. cholerae O1/O139 whole cell cholera vaccine: Comparison with other mucosal and systemic responses.
   
   Shamsuzzaman S, Ahmed T, Mannoor K, Begum YA, Bardhan PK, Sack RB, Sack DA, Svennerholm AM, Holmgren J, Qadri F.
   International Centre for Diarrhoeal Disease Research, Bangladesh, GPO Box 128, Dhaka 1000, Bangladesh; Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka 1000, Bangladesh.
   
   The emergence of V. cholerae O139 serogroup of V. cholerae capable of causing severe dehydrating cholera has over the decade led to efforts in formulation of vaccines to protect against this pathogen. Although the prevalence of diarrhea due to V. cholerae O139 has recorded a decrease, efforts on vaccine development continues to formulate an oral vaccine capable of stimulating the gut mucosal system. We have studied the mucosal immunogenicity in Bangladeshi adults to a killed whole cell (WC) bivalent cholera vaccine composed of V. cholerae O139 as well as V. cholerae O1 strains together with the recombinant cholera toxin B subunit (CTB) (WC-O1/O139/CTB) and compared the immune responses to that obtained with the licensed monovalent cholera vaccine, Dukoral (WC-O1/CTB). Direct estimation of the WC-O1/O139/CTB vaccine-specific mucosal responses were carried out using lymphocytes isolated from duodenal biopsies, intestinal lavage fluid and feces. The vaccine induced robust antibody-secreting cell responses in the duodenum specific to CTB as well as the O1 and O139 lipopolysaccharide (LPS). Magnitude of response was higher in the gut than in the circulation in all three antibody isotypes. The CTB and LPS-specific mucosal antibody responses were also seen in intestinal lavage fluid and fecal extracts. Vibriocidal antibody responses in plasma were observed to both the V. cholerae O1 and O139 serogroups (76% and 57% response rates, respectively). Plasma IgA and IgG responses to CTB and IgA responses to both O1 and O139 LPS were elevated. The immune responses were comparable to that seen to the monovalent WC-O1/CTB recipients in all components studied. Overall, the bivalent cholera vaccine induces strong mucosal responses and the addition of the O139 component does not interfere with the responses to the licensed vaccine Dukoral. This sets the ground for testing such vaccines in large field trials in Bangladesh and also demonstrates that addition of other vibrio components to the existing cholera vaccine does not alter the responses to the O1 vaccine components.
   PMID: 19146897
   
   www.ncbi.nlm.nih.gov/pubmed/19146897?
10. flosz 18 januari 2009 12:59

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    Int J Med Microbiol. 2008 Dec 31
    Genetic stability of vaccine strain Salmonella Typhi Ty21a over 25 years.
    
    Kopecko DJ, Sieber H, Ures JA, Fürer A, Schlup J, Knof U, Collioud A, Xu D, Colburn K, Dietrich G.
    Laboratory of Enteric and Sexually Transmitted Diseases, Center for Biologics Evaluation and Research, Food and Drug Administration, 29 Lincoln Drive, NIH Campus, Bldg. 29/420, HFM440, Bethesda, MD 20892, USA.
    
    The attenuated live bacterial vaccine strain Salmonella enterica Serovar Typhi Ty21a is the main constituent of Vivotif, the only licensed oral vaccine against typhoid fever. The strain was developed in the 1970s by chemical mutagenesis. In the course of this mutagenesis, a number of mutations were introduced into the vaccine strain. Characterisation of the vaccine strain during development as well as release of master- and working seed lots (MSL and WSL) and commercial batches is based on phenotypic assays assessing microbiological and biochemical characteristics of Ty21a. In the current study, we have analysed by DNA sequencing the specific mutations originally correlated with the attenuation of strain Ty21a. These data demonstrate the stability of these mutations for MSLs and WSLs of Ty21a produced between 1980 and 2005. Finally, we have confirmed the correlation of these genetic mutations with the expected phenotypic attenuations for the seed lots used in vaccine manufacture over 25 years.
    PMID: 19121604
    www.ncbi.nlm.nih.gov/pubmed/19121604?
    
11. [verwijderd] 28 januari 2009 17:18

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    Sensitive and Specific Detection of the Non-Human Sialic Acid N-Glycolylneuraminic Acid In Human Tissues and Biotherapeutic Products
    
    Conclusions
    We report a reliable antibody-based method for highly sensitive and specific detection of the non-human sialic acid Neu5Gc in human tissues and biotherapeutic products that has not been previously described.
    
    www.pubmedcentral.nih.gov/articlerend...
    
    Iemand met meer biotech achtergrond die dit helderder kan maken?
12. [verwijderd] 8 februari 2009 19:55

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    Patrys weer met Percivia !
    
    High level expression of functional human IgMs in human PER.C6® cells
    Anna Tchoudakova, Frank Hensel, Alec Murillo, Bernie Eng, Marketa Foley, Lakee Smith, Frank Schoenen, Antonia Hildebrand, Arndt-René Kelter, Leodevico L. Ilag, H. Peter Vollmers, Stephanie Brandlein, Jane McIninch, John Chon, Gene Lee and Marco Cacciuttolo
    
    volume 1 | issue 2
    March/April 2009
    
    Natural IgM antibodies play an important role in the body’s defense mechanisms against transformed cells in the human body and are currently being exploited both in prognoses of malignant lesions and in the therapy of cancer patients. However, despite growing interest and clinical promise, thus far the IgM class of antibodies has failed to gain widespread commercial interest as these are considered to be difficult to produce recombinantly. IgMs are polymeric and have a relatively large mass. In addition, IgM molecules are heavily glycosylated and, when produced in non-human cell lines, they may contain non-human glycan structures which may be potentially immunogenic. Clearly, production systems capable of expressing human recombinant IgM antibodies are needed. We have successfully used PER.C6® cells – a human cell line - to generate three separate human recombinant monoclonal IgMs in suspension cultures in protein-free medium. All three of the IgMs were constructed with joining (J) chain and were expressed in the pentameric form. One of the IgMs was also expressed as a hexamer without J chain. Clones with cell specific productivities greater than 20 pg/cell/day were generated, which led to yields of 0.5 g/L to 2g/L in fed-batch production. All the IgMs expressed were biologically active as shown in binding and cytotoxicity assays. These studies demonstrate the potential of PER.C6® cells for the production of high levels of functional recombinant IgM and other polymeric molecules, using a straightforward and rapid stable cell line generation method.
    
    www.landesbioscience.com/journals/mab...
13. flosz 8 februari 2009 20:44

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    quote:

    gocrucellgo schreef:

    Patrys weer met Percivia !
    

    www.youtube.com/watch?v=VWf1MdHv80Q go!!!
    
    Cell Line Generation
    The PER.C6® cell line development platform is designed to be user-friendly and robust, with rapid development timelines from transfection to stable cell line. All steps are performed in serum-free, chemically defined medium.
    • Suspension-adapted host cells – no need to adapt to suspension culture during cell line development
    • Hundreds of clones screened, using standard mammalian cell line development strategies
    • Rapid development timelines – transfection to Research Cell Bank in 5 months
    • High productivities, without amplification – cell lines with cell specific productivities of >50 pg/cell/day!
    • PERCIVIA offers a full suite of cell line development support including vector construction, serum-free cell line generation up to Research Cell Bank, and cell line stability studies (>50 generations in culture, in the presence and absence of selective pressure). Research Cell Banks are tested for mycoplasma and sterility. A cGMP Master Cell Banking service is also available, through PERCIVIA’s partnership with DSM Biologics. In addition, full support and hands-on training on the use of PER.C6® cells for cell line development is available to all licensees.
    
    Cell Culture: Batch and Fed-Batch Production Process
    One of the key advantages of the PER.C6® technology is the high performance of its platform production process. An ongoing area of development at PERCIVIA is in the continuing improvement of this baseline batch and fed-batch processes for material generation in support of pre-clinical and clinical research.
    Some of the key features of the platform batch and fed-batch production processes include:
    • High performance – PER.C6® cells may be cultured to very high concentrations in batch and fed-batch cultures. Viable cell concentration (Xv) > 12 X 106 cells/mL in batch and > 25 X 106 cells/mL in fed-batch are typically achieved (Figure 1). With these high cell concentrations, integral viable cell (IVC) of over 100 billion cell days per liter (bcdl) in batch and over 300 bcdl in fed-batch systems are routinely achieved. This translates to productivities of ~ 2 g/L in batch and ~ 8 g/L in fed-batch with clones whose productivities average less than 30 picograms/cell/day (pcd)
    • Safety – all the media and supplements used in batch and fed-batch processes are animal-derived component free and chemically defined.
    • Easy implementation – the processes are designed to be easy to implement in most mammalian cell culture labs and manufacturing plants. The processes are simplified to require minimal operator intervention and should be easily recognizable to scientists and operators familiar with standard mammalian cell culture techniques and practices
    
    XD™ Process
    The XD™ process takes advantage of the PER.C6® cell’s inherent ability to withstand high shear and grow to very high concentrations. This process uses Refine’s Alternating Tangential Flow (ATF) system to perfuse fresh medium through the bioreactor and remove metabolic by-products while retaining the cells. The result is a process in which PER.C6® cells may be cultured to extreme densities and very high product concentration.
    The features of the XD™ process include the following:
    • Extreme cell concentrations – by continuously perfusing in fresh nutrients and removing metabolic by-products, PER.C6® cells are kept growing exponentially throughout the process. Typically viable cell concentration of 100 – 150 X 106 cells/mL is normally achieved in ~ 2 weeks
    • Extreme productivity – Typical process IVC is ~ 500 bcdl in two weeks. This means, with a cell line of typical productivity (30 pcd), product concentration of10 - 15 g/L may be routinely achieved
    • Consistency – continuous perfusion of fresh medium and removal of metabolic byproducts means the cells are kept in a relatively constant environment throughout the process. This results in more consistent product quality profile and culture performance may be easily predicted using a relatively straight-forward simulation model
    Protein Purification
    Downstream process development is an integral part of the PER.C6® platform. Its focus is the use of economic and scalable technologies that enable the utilization of the full potential for this cell line and result high quality protein. PERCIVIA’s DSP philosophy is to creatively merge established technologies as well as breakthrough ones such as single use membrane chromatography techniques
    • Design robust, cost effective, scalable platform processes for the manufacturing of Mabs and other recombinant molecules
    • Accommodate high titer (> 5 g/L), and high cell density (> 30 million cells/mL) cultures
    • Effective and scalable methods to harvest extreme cell density cultures (> 100 million cells/mL)
    • Incorporates high capacity chromatography media (> 90 mg/L DBC for Mabs) and other breakthrough technologies
    www.percivia.com/researchDevt/index.html
    

    Bijlage:
14. flosz 10 februari 2009 21:28

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    • flosz

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    Daar is "oud-Cruceller" Menzo:
    
    Cancer Gene Therapy advance online publication 30 January 2009; doi: 10.1038/cgt.2009.4
    
    Fiber-chimeric adenoviruses expressing fibers from serotype 16 and 50 improve gene transfer to human pancreatic adenocarcinoma
    
    K F D Kuhlmann1, M A van Geer2, C T Bakker2, J E M Dekker2, M J E Havenga3, R P J Oude Elferink2, D J Gouma1, P J Bosma2 and J G Wesseling2,4
    1. 1Department of Surgery, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
    2. 2Liver Center, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
    3. 3TNO Biosciences, Leiden, The Netherlands
    Correspondence: Dr PJ Bosma, AMC Liver Center, Academic Medical Center, University of Amsterdam, Meibergdreef 69/71, 1105 BK Amsterdam, The Netherlands. E-mail: p.j.bosma@amc.uva.nl
    4Current address: Corning BV, Schiphol-Rijk, The Netherlands.
    
    Received 16 July 2008; Revised 8 October 2008; Accepted 20 November 2008; Published online 30 January 2009
    Survival of patients with pancreatic cancer is poor. Adenoviral (Ad) gene therapy employing the commonly used serotype 5 reveals limited transduction efficiency due to the low amount of coxsackie-adenovirus receptor on pancreatic cancer cells. To identify fiber-chimeric adenoviruses with improved gene transfer, a library of Ad vectors based on Ad5 and carrying fiber molecules consisting of 16 other serotypes were transduced to human pancreatic carcinoma cell lines. Adenoviruses containing fibers from serotype 16 and 50 showed increased gene transfer and were further analyzed. In a gene-directed prodrug activation system using cytosine deaminase, these adenoviruses proved to be effective in eradicating primary pancreatic tumor cells. Fiber-chimeric Ad5 containing fiber 16 and wild-type Ad5 were also transduced ex vivo to slices of normal human pancreatic tissue and pancreatic carcinoma tissue obtained during surgery. It was shown that fiber-chimeric Ad5 with fiber 16 revealed an improved gene delivery to primary pancreatic tumor tissue compared to Ad5. In conclusion, fiber-chimeric adenoviruses carrying fiber 16 and 50 reveal a significantly enhanced gene transfer and an increased specificity to human pancreatic adenocarcinoma compared to Ad5, whereas transduction to normal pancreatic tissue was decreased. These findings expand the therapeutic window of Ad gene therapy for pancreatic cancer.
    We thank Jerome Custers (Crucell BV, Leiden) for providing the various virus batches.
    www.nature.com/cgt/journal/vaop/ncurr...
    
15. flosz 11 februari 2009 19:48

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    Vaccine. 2009 Feb 4.
    Recombinant measles viruses expressing single or multiple antigens of human immunodeficiency virus (HIV-1) induce cellular and humoral immune responses.
    
    Liniger M, Zuniga A, Morin TN, Combardiere B, Marty R, Wiegand M, Ilter O, Knuchel M, Naim HY.
    Crucell - Berna Biotech LTD, Rehhagstrasse 79, 3018 Bern, Switzerland.
    
    Recombinant measles viruses (rMV) based on the live attenuated measles vaccine strain (MVb) expressing antigens of HIV-1 clade B were generated by reverse genetics. Recombinants expressing single or double antigens of HIV-1 (rMV-HIV) were genetically highly stable on human diploid cells. The production process of these viruses was essentially similar to the parental MV strain, yielding comparative end titers. Immunization of tg-mice by different regimens and formulations showed potent humoral and cellular immune responses against MV and HIV antigens. Recombinant MV-HIV expressing Gag protein conferred protective immunity in tg-mice after a high-dose pseudochallenge with recombinant vaccinia virus. In addition, rMV-HIV boosted anti-HIV antibodies, in the presence of pre-existing anti-vector antibodies.
    PMID: 19200842
    www.ncbi.nlm.nih.gov/pubmed/19200842
    *********************************
    
    Vaccine. 2009 Feb 4
    Economic benefits for the family of inactivated subunit virosomal influenza vaccination of healthy children aged 3-14 years during the annual health examination in private paediatric offices.
    
    Salleras L, Navas E, Domínguez A, Ibáñez D, Prat A, Garrido P, Asenjo MA, Torner N.
    Department of Public Health, School of Medicine, University of Barcelona, Spain; Preventive Medicine Unit, Hospital Clinic Barcelona, Spain.
    
    Taking the results of a prospective cohort study by our group that evaluated the effectiveness of the inactivated subunit virosomal influenza vaccine (Inflexal V((R)), Crucell-Berna) in the prevention of influenza-related diseases and the reduction of its negative economic consequences, the economic costs and benefits for the family of vaccinating a theoretical cohort of 1000 healthy children aged 3-14 years with no risk factors with one dose of vaccine during the yearly health examination were quantiified. The economic analysis was carried out from the family perspective and the time horizon of the study was established at 6 months. In the base case, the net present value was 21,551.62 euros (21.5 euros per vaccinated child), and the benefit-cost ratio was 2.15, meaning that 1.15 euros is saved per euro invested.
    PMID: 19200830
    www.ncbi.nlm.nih.gov/pubmed/19200830
    
16. flosz 11 februari 2009 19:58

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    Vaccine. 2009 Feb 4.
    Flu vaccination with a virosomal vaccine does not affect clinical course and immunological parameters in scleroderma patients.
    
    Setti M, Fenoglio D, Ansaldi F, Filaci G, Bacilieri S, Sticchi L, Ferrera A, Indiveri F, Ghio M.
    Clinic of Internal Medicine and Clinical Immunology, Department of Internal Medicine (DIMI), San Martino Hospital, University of Genoa, Viale Benedetto XV 6, 16153 Genoa, Italy.
    
    Safety and efficacy of adjuvanted vaccines in autoimmune individuals raises growing clinical and scientific interest. Protection from influenza would bring particular benefits in these patients with common cardiac and respiratory impairment. This study evaluates efficacy, clinical safety and immune effects of the administration of a single dose of a virosomal flu vaccine in 46 scleroderma patients. The following parameters were evaluated before and after administration of Inflexal: clinical conditions, inflammation and autoimmunity parameters, humoral response, lymphocyte proliferation and cytokine production upon flu antigen stimulation by specific and non-specific cells. Inflexal was found effective in scleroderma patients. In no subject was worsening of clinical conditions, inflammation and immunological parameters observed.
    PMID: 19200840
    www.ncbi.nlm.nih.gov/pubmed/19200840
    **********************
    
    Immunol Invest. 2009;38(1):67-75.
    A comparison between ((3)H)-thymidine incorporation and isothermal microcalorimetry for the assessment of antigen-induced lymphocyte proliferation.
    
    Murigande C, Regenass S, Wirz D, Daniels AU, Tyndall A.
    Department of Rheumatology, Felix Platter Spital, Basel, Switzerland. claire.murigande@unibas.ch
    
    Lymphocyte transformation tests (LTT) are time-consuming radioactive assays used in the clinic for the determination of allergic drug reactions and extensively in basic immunological research. In the present study we propose an alternative method in the monitoring of T-cell responses by isothermal microcalorimetric (IMC) measurements of overall cellular heat production as a function of time. For mitogen-induced lymphocyte proliferation, we analyzed a concentration dependent effect of phytohemaglutinin (PHA) and both tests showed a good correlation. This was also the case for specific antigenic stimulation with Varidase(R) or tetanus toxoid. On the other hand, antigen-induced lymphocyte proliferation analyzed by pre and post influenza vaccine (Inflexal(R) V) samples, showed no such correlation. Our study suggests that IMC measurements, despite the advantages of simplicity, on-line recording of metabolic activity and no use of radioactivity, may be limited to monitoring mitogen-induced lymphocyte proliferation.
    PMID: 19172486
    www.ncbi.nlm.nih.gov/pubmed/19172486
    
    
17. flosz 17 maart 2009 13:04

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    J Gen Virol. 2009 Mar 12.
    
    Adenovirus Serotype 5 Infects Human Dendritic Cells via a CAR-Independent Receptor Pathway Mediated by Lactoferrin and DC-SIGN.
    Adams WC, Bond E, Havenga MJ, Holterman L, Goudsmit J, Karlsson-Hedestam GB, Koup RA, Loré K.
    Karolinska Institutet;
    
    The coxsackievirus and adenovirus receptor (CAR) is the described primary receptor for Adenovirus serotype 5 (Ad5), a common human pathogen that has been exploited as a viral vector for gene therapy and vaccination. Here, we report that monocytes and dendritic cells (DCs) such as freshly isolated human blood myeloid DCs, plasmacytoid DCs, and monocyte-derived DCs are susceptible to recombinant Ad5 (rAd5) infection despite their lack of CAR expression. Langerhans cells and dermal DCs from skin express CAR but blocking CAR only partly decreased rAd5 infection, together suggesting that other receptor pathways mediate viral entry of these cells. Lactoferrin (Lf), an abundant protein in many bodily fluids known for its anti-viral and anti-bacterial properties, promoted rAd5 infection in all cell populations except plasmacytoid DCs using a CAR-independent process. Lf phenotypically differentiated the DCs but cell activation only played a minor role in the increase in infection frequencies. The C-type lectin receptor DC-SIGN facilitated viral entry of rAd5-Lf complexes and this was dependent on high-mannose type N-linked glycans on Lf. We therefore speculate that Lf present at high levels at mucosal sites can facilitate rAd5 attachment and enhance infection of DCs. A better understanding of the tropism and receptor mechanisms of Ad5 can help explain Ad5 pathogenesis and guide the engineering of improved rAd vectors.
    PMID: 19282435
    www.ncbi.nlm.nih.gov/pubmed/19282435
    
18. [verwijderd] 8 april 2009 18:47

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    Even wat luchtigs tussendoor na al die serieuze postings.
    
    Suitability of PER.C6® cells to generate epidemic and pandemic influenza vaccine strains by reverse genetics
    
    W Koudstaal, L Hartgroves, M Havenga, I Legastelois, C Ophorst, M Sieuwerts, D Zuijdgeest, R Vogels, J Custers, E de Boer-Luijtze, O de Leeuw, L Cornelissen, J Goudsmit and W Barclay
    
    Reverse genetics, the generation of influenza viruses from cDNA, presents a rapid method for creating vaccine strains. The technique necessitates the use of cultured cells. Due to technical and regulatory requirements, the choice of cell lines for production of human influenza vaccines is limited. PER.C6® cells, among the most extensively characterized and documented cells, support growth of all influenza viruses tested to date, and can be grown to high densities in large bioreactors in the absence of serum or micro carriers. Here, the suitability of these cells for the generation of influenza viruses by reverse genetics was investigated. A range of viruses reflective of vaccine strains was rescued exclusively using PER.C6 cells by various transfection methods, including an animal component-free procedure. Furthermore, a whole inactivated vaccine carrying the HA and NA segments of A/HK/156/97 (H5N1) that was both rescued from and propagated on PER.C6 cells, conferred protection in a mouse model. Thus PER.C6 cells provide an attractive platform for generation of influenza vaccine strains via reverse genetics.
    
    pubget.com/search?q=Vaccine%5Blatest%...
19. flosz 8 april 2009 19:18

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    Haha, de leukste van vandaag!
    
    Vaccine, Volume 27, Issue 19, 28 April 2009, Pages 2588-2593
    
    W. Koudstaala, 1, , , L. Hartgrovesb, 1, M. Havengac, I. Legasteloisd, C. Ophorsta, M. Sieuwertsa, D. Zuijdgeesta, R. Vogelsa, J. Custersa, E. de Boer-Luijtzee, O. de Leeuwe, L. Cornelissene, J. Goudsmita and W. Barclayb
    
    aCrucell Holland BV, Leiden, The Netherlands
    
    bDepartment of Virology, Imperial College, St. Mary's Campus, London, United Kingdom
    
    cTNO Biosciences, Leiden, The Netherlands
    
    dSanofi-Pasteur, Lyon, France
    
    eCentral Veterinary Institute, Wageningen UR, Lelystad, The Netherlands
    
    Received 21 October 2008; revised 2 February 2009; accepted 11 February 2009. Available online 20 February 2009.
20. nelis h 8 april 2009 20:01

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    de link erbij: www.sciencedirect.com/science?_ob=Art...