{"title":"Streamline Your Cell Line Screening: With the Automated, Microscale, Stirred Tank, Single-Use Bioreactor","authors":"T. Ward","doi":"10.12665/J134.WARD","DOIUrl":"https://doi.org/10.12665/J134.WARD","url":null,"abstract":"","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"13 1","pages":"60-61"},"PeriodicalIF":0.0,"publicationDate":"2015-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2015-01-16DOI: 10.12665/J134.NitzkiHerbig
F. Nitzki, Elmar Herbig
Principle of In Situ Hybridization ISH represents an alternative to immunohistochemical staining if adequate antibodies are not available, and is employed in diverse areas of research. This type of hybridization uses specific nucleic acid fragments (probes) that are complementary to the target sequence in order to detect specific transcripts. Such probes may consist of DNA or of RNA. Meanwhile, RNA probes are frequently utilized. Such probes are commonly labeled with the molecule digoxigenin (DIG), which normally occurs in the plant species Digitalis purpurea. DIG labeling enables the specifically bound probes in a tissue section to be visualized by enzyme-conjugated anti-DIG antibodies. For this purpose, after incubation of tissue sections with antibody, the appropriate substrate is pipetted onto the sections and converted by an immunoenzymatic reaction into a visible colorant (Figure 1). This method can be used to analyze the activity of specific genes for research projects or diagnostic procedures. Further practical information on ISH is provided in Wilcox‘s overview article.[2] This article discusses the results of the ISH that was carried out as part of a cancer research project. The analyzed skin samples were obtained from genetically modified mice. Based on targeted manipulation of the tumor suppressor gene Patched (Ptch) by homozygous knockout, the Ptch-knockout mice developed basal cell carcinomas.[3] These skin tumors are the ones that most commonly occur in humans. They frequently exhibit increased activity of the signaling pathway that is regulated by Ptch. Inactivation of this important component in this mouse model pathologically activates the signaling pathway. This results in an increased expression of the target gene Gli1 — a transcription factor that activates a variety of other genes — in tumor cells, and expression of Gli1 can be detected using ISH.
{"title":"In Situ Hybridization: The Importance of Ultrapure Water for RNA Technologies","authors":"F. Nitzki, Elmar Herbig","doi":"10.12665/J134.NitzkiHerbig","DOIUrl":"https://doi.org/10.12665/J134.NitzkiHerbig","url":null,"abstract":"Principle of In Situ Hybridization ISH represents an alternative to immunohistochemical staining if adequate antibodies are not available, and is employed in diverse areas of research. This type of hybridization uses specific nucleic acid fragments (probes) that are complementary to the target sequence in order to detect specific transcripts. Such probes may consist of DNA or of RNA. Meanwhile, RNA probes are frequently utilized. Such probes are commonly labeled with the molecule digoxigenin (DIG), which normally occurs in the plant species Digitalis purpurea. DIG labeling enables the specifically bound probes in a tissue section to be visualized by enzyme-conjugated anti-DIG antibodies. For this purpose, after incubation of tissue sections with antibody, the appropriate substrate is pipetted onto the sections and converted by an immunoenzymatic reaction into a visible colorant (Figure 1). This method can be used to analyze the activity of specific genes for research projects or diagnostic procedures. Further practical information on ISH is provided in Wilcox‘s overview article.[2] This article discusses the results of the ISH that was carried out as part of a cancer research project. The analyzed skin samples were obtained from genetically modified mice. Based on targeted manipulation of the tumor suppressor gene Patched (Ptch) by homozygous knockout, the Ptch-knockout mice developed basal cell carcinomas.[3] These skin tumors are the ones that most commonly occur in humans. They frequently exhibit increased activity of the signaling pathway that is regulated by Ptch. Inactivation of this important component in this mouse model pathologically activates the signaling pathway. This results in an increased expression of the target gene Gli1 — a transcription factor that activates a variety of other genes — in tumor cells, and expression of Gli1 can be detected using ISH.","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"6 1","pages":"56-59"},"PeriodicalIF":0.0,"publicationDate":"2015-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229633","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Use of Interactive Laser-Scanning Imaging Cytometry (ILIC) for Real-Time Cytotoxicity Assessment of Bacteriocins Against Cultured Mammalian Cells","authors":"S. Murinda, R. Roberts, E. Kunze, K. Rashid","doi":"10.12665/J134.MURINDA","DOIUrl":"https://doi.org/10.12665/J134.MURINDA","url":null,"abstract":"","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"13 1","pages":"21-32"},"PeriodicalIF":0.0,"publicationDate":"2015-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229618","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Production of Kojic Acid by Aerobic Aspergillus Fermentation","authors":"S. Lakhawat, Jignesh Chaudhary, A. Pathak","doi":"10.12665/J133.LAKHAWAT","DOIUrl":"https://doi.org/10.12665/J133.LAKHAWAT","url":null,"abstract":"","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"13 1","pages":"62-69"},"PeriodicalIF":0.0,"publicationDate":"2014-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229449","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Contract Pharma Manufacturer Optimizes Quality Check Process With the Cubis Lab Balance","authors":"Matthias Ude","doi":"10.12665/J133.UDE","DOIUrl":"https://doi.org/10.12665/J133.UDE","url":null,"abstract":"","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"13 1","pages":"61-61"},"PeriodicalIF":0.0,"publicationDate":"2014-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229518","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A n increasing number of clinical trials, and the recent approval of the first gene therapy in Europe, alipogene tiparvovec (Glybera®, uniQure), holds promise for recombinant adeno-associated virus (rAAV) to become a mainstay in clinical practice. Since the molecular cloning of AAV in the 1980s, a plethora of production protocols/manufacturing systems for generating rAAV vectors have been developed. uniQure’s manufacturing platform, which has received validation through regulatory approval, is also capable of supporting industrial-scale production based on the baculovirus expression vector system (BEVS) and insect cells. In this paper, we review the molecular process optimization of the various components of uniQure’s rAAV production platform.
{"title":"Insect Cell-Based Recombinant Adeno-Associated Virus Production: Molecular Process Optimization","authors":"J. Lubelski, W. Hermens, H. Petry","doi":"10.12665/J133.LUBELSKI","DOIUrl":"https://doi.org/10.12665/J133.LUBELSKI","url":null,"abstract":"A n increasing number of clinical trials, and the recent approval of the first gene therapy in Europe, alipogene tiparvovec (Glybera®, uniQure), holds promise for recombinant adeno-associated virus (rAAV) to become a mainstay in clinical practice. Since the molecular cloning of AAV in the 1980s, a plethora of production protocols/manufacturing systems for generating rAAV vectors have been developed. uniQure’s manufacturing platform, which has received validation through regulatory approval, is also capable of supporting industrial-scale production based on the baculovirus expression vector system (BEVS) and insect cells. In this paper, we review the molecular process optimization of the various components of uniQure’s rAAV production platform.","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"13 1","pages":"6-11"},"PeriodicalIF":0.0,"publicationDate":"2014-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Failure Mode and Effect Analysis (FMEA) as a Quality by Design (QbD) Tool for Managing Biopharmaceutical Product Development and Manufacturing Risks","authors":"Mark Witcher","doi":"10.12665/J133.WITCHER","DOIUrl":"https://doi.org/10.12665/J133.WITCHER","url":null,"abstract":"","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"13 1","pages":"39-46"},"PeriodicalIF":0.0,"publicationDate":"2014-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229562","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2014-10-13DOI: 10.12665/J133.BOTTOVALEE
Iveta Bottová, Lena Lee
{"title":"Validation Study of the Vi-CELL XR for Dendritic Cell Counting","authors":"Iveta Bottová, Lena Lee","doi":"10.12665/J133.BOTTOVALEE","DOIUrl":"https://doi.org/10.12665/J133.BOTTOVALEE","url":null,"abstract":"","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"13 1","pages":"32-37"},"PeriodicalIF":0.0,"publicationDate":"2014-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Which Factors to Consider When Selecting an Analytical Method for Cell Culture Fermentation: A Comparison of Four Different Metabolic Analyzer Instruments","authors":"C. Kurtz, J. Smelko","doi":"10.12665/J133.KURTZ","DOIUrl":"https://doi.org/10.12665/J133.KURTZ","url":null,"abstract":"","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"13 1","pages":"12-31"},"PeriodicalIF":0.0,"publicationDate":"2014-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Aguilera, L. Muñoz, Y. Bermúdez, Danae Arias, Yamila Martínez, Gerardo García, L. Hernández, E. Martínez, R. Valdés
{"title":"Validation of a Chromogenic Substrate Method for Biological Activity Quantification of Streptokinase","authors":"A. Aguilera, L. Muñoz, Y. Bermúdez, Danae Arias, Yamila Martínez, Gerardo García, L. Hernández, E. Martínez, R. Valdés","doi":"10.12665/J133.VALDES","DOIUrl":"https://doi.org/10.12665/J133.VALDES","url":null,"abstract":"","PeriodicalId":88836,"journal":{"name":"Bioprocessing","volume":"257 1","pages":"49-59"},"PeriodicalIF":0.0,"publicationDate":"2014-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66229546","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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