Robert M Kotin

Summary

Affiliation: National Institutes of Health
Country: USA

Publications

  1. pmc Strategies for manufacturing recombinant adeno-associated virus vectors for gene therapy applications exploiting baculovirus technology
    Alejandro Negrete
    Birmingham University, UK
    Brief Funct Genomic Proteomic 7:303-11. 2008
  2. doi request reprint Large-scale production of recombinant adeno-associated viral vectors
    Alejandro Negrete
    Laboratory of Biochemical Genetics, National Heart, Lung, Blood Institute, Bethesda, MD, USA
    Methods Mol Biol 433:79-96. 2008
  3. ncbi request reprint Serum-free production and column purification of adeno-associated virus type 5
    Richard H Smith
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, Building 10, Room 7N 264, 9000 Rockville Pike, Bethesda, MD 20892, USA
    J Virol Methods 114:115-24. 2003
  4. ncbi request reprint Process optimization of large-scale production of recombinant adeno-associated vectors using dielectric spectroscopy
    Alejandro Negrete
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, US National Institutes of Health, 10 Center Drive, NIH, Building 10, Room 7D05, Bethesda, MD 20892, USA
    Appl Microbiol Biotechnol 76:761-72. 2007
  5. pmc A simplified baculovirus-AAV expression vector system coupled with one-step affinity purification yields high-titer rAAV stocks from insect cells
    Richard H Smith
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, Bethesda, Maryland 20892, USA
    Mol Ther 17:1888-96. 2009
  6. doi request reprint Chromatography-based purification of adeno-associated virus
    Richard H Smith
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD, USA
    Methods Mol Biol 434:37-54. 2008
  7. pmc Reproducible high yields of recombinant adeno-associated virus produced using invertebrate cells in 0.02- to 200-liter cultures
    Sylvain Cecchini
    Laboratory of Molecular Virology and Gene Therapy, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
    Hum Gene Ther 22:1021-30. 2011
  8. ncbi request reprint Economized large-scale production of high yield of rAAV for gene therapy applications exploiting baculovirus expression system
    Alejandro Negrete
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, US National Institutes of Health, Bethesda, MD 20892, USA
    J Gene Med 9:938-48. 2007
  9. pmc Large-scale recombinant adeno-associated virus production
    Robert M Kotin
    Laboratory of Molecular Virology and Gene Therapy, Center for Developmental Biology and Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
    Hum Mol Genet 20:R2-6. 2011
  10. pmc Production of recombinant adeno-associated vectors using two bioreactor configurations at different scales
    Alejandro Negrete
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, US National Institutes of Health, Bethesda, MD 20892, USA
    J Virol Methods 145:155-61. 2007

Collaborators

Detail Information

Publications30

  1. pmc Strategies for manufacturing recombinant adeno-associated virus vectors for gene therapy applications exploiting baculovirus technology
    Alejandro Negrete
    Birmingham University, UK
    Brief Funct Genomic Proteomic 7:303-11. 2008
    ..The yields obtained with these optimized processes exceed approximately 1 x 10(14) vector particles per liter of cell culture suitable for pre-clinical and clinical trials and possible commercialization...
  2. doi request reprint Large-scale production of recombinant adeno-associated viral vectors
    Alejandro Negrete
    Laboratory of Biochemical Genetics, National Heart, Lung, Blood Institute, Bethesda, MD, USA
    Methods Mol Biol 433:79-96. 2008
    ..This represents significant progress toward establishing a robust large-scale process at industry level...
  3. ncbi request reprint Serum-free production and column purification of adeno-associated virus type 5
    Richard H Smith
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, Building 10, Room 7N 264, 9000 Rockville Pike, Bethesda, MD 20892, USA
    J Virol Methods 114:115-24. 2003
    ..The method described in this report does not require partial purification by density centrifugation, detergent treatment, or solvent extraction to achieve efficient levels of column binding and vector purification...
  4. ncbi request reprint Process optimization of large-scale production of recombinant adeno-associated vectors using dielectric spectroscopy
    Alejandro Negrete
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, US National Institutes of Health, 10 Center Drive, NIH, Building 10, Room 7D05, Bethesda, MD 20892, USA
    Appl Microbiol Biotechnol 76:761-72. 2007
    ..As part of large-scale optimization studies, this new finding will facilitate the bioprocessing scale-up of rAAV and other bioproducts...
  5. pmc A simplified baculovirus-AAV expression vector system coupled with one-step affinity purification yields high-titer rAAV stocks from insect cells
    Richard H Smith
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, Bethesda, Maryland 20892, USA
    Mol Ther 17:1888-96. 2009
    ..Using the described method, we obtained an average yield of 7 x 10(4) purified rAAV particles per cell (range: 3.7 x 10(4) to 9.6 x 10(4)) from suspension cultures of recombinant baculovirus-infected insect cells...
  6. doi request reprint Chromatography-based purification of adeno-associated virus
    Richard H Smith
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD, USA
    Methods Mol Biol 434:37-54. 2008
    ..This method provides a basic strategy, or "platform," that can be adapted to the purification of other recombinant AAV vector serotypes...
  7. pmc Reproducible high yields of recombinant adeno-associated virus produced using invertebrate cells in 0.02- to 200-liter cultures
    Sylvain Cecchini
    Laboratory of Molecular Virology and Gene Therapy, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
    Hum Gene Ther 22:1021-30. 2011
    ..The ease and reproducibility of rAAV production in 200-liter bioreactors suggest that the limit has not been reached, and 500-liter productions are planned...
  8. ncbi request reprint Economized large-scale production of high yield of rAAV for gene therapy applications exploiting baculovirus expression system
    Alejandro Negrete
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, US National Institutes of Health, Bethesda, MD 20892, USA
    J Gene Med 9:938-48. 2007
    ..However, production in larger volumes is limited by the stability of the BEVs and amount of BEVs needed to achieve the target MOI of 3 per BEV. Here, the production parameters were optimized and the baculovirus stability was determined...
  9. pmc Large-scale recombinant adeno-associated virus production
    Robert M Kotin
    Laboratory of Molecular Virology and Gene Therapy, Center for Developmental Biology and Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
    Hum Mol Genet 20:R2-6. 2011
    ..Issues relating to developing new and improving existing rAAV production methods are discussed...
  10. pmc Production of recombinant adeno-associated vectors using two bioreactor configurations at different scales
    Alejandro Negrete
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, US National Institutes of Health, Bethesda, MD 20892, USA
    J Virol Methods 145:155-61. 2007
    ..These results demonstrate that producing rAAV in large scale using BEVs is reproducible, scalable, and independent of the bioreactor configuration...
  11. pmc Evidence of prior exposure to human bocavirus as determined by a retrospective serological study of 404 serum samples from adults in the United States
    Sylvain Cecchini
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA
    Clin Vaccine Immunol 16:597-604. 2009
    ..Using ELISA, we tested 404 human serum samples and established a range of antibody titers in a large U.S. adult population sample...
  12. pmc Producing recombinant adeno-associated virus in foster cells: overcoming production limitations using a baculovirus-insect cell expression strategy
    Tamas Virag
    Molecular Virology and Gene Delivery Section, Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
    Hum Gene Ther 20:807-17. 2009
    ....
  13. ncbi request reprint Structural unity among viral origin binding proteins: crystal structure of the nuclease domain of adeno-associated virus Rep
    Alison Burgess Hickman
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Cell 10:327-37. 2002
    ..The active site, located at the bottom of a positively charged cleft, is formed by the spatial convergence of a divalent metal ion and two conserved sequence motifs that define the rolling circle replication superfamily...
  14. ncbi request reprint The DNA minor groove binding agents Hoechst 33258 and 33342 enhance recombinant adeno-associated virus (rAAV) transgene expression
    Lina Li
    Laboratory of Biochemical Genetics, NHLBI, National Institutes of Health, Bethesda, MD 20892, USA
    J Gene Med 7:420-31. 2005
    ..Recombinant adeno-associated viruses (rAAV) are commonly used in pre-clinical and clinical gene transfer studies. However, the relatively slow kinetics of rAAV transgene expression complicates in vitro and in vivo experiments...
  15. pmc Production and characterization of novel recombinant adeno-associated virus replicative-form genomes: a eukaryotic source of DNA for gene transfer
    Lina Li
    Laboratory of Molecular Virology and Gene Therapy, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, United States of America
    PLoS ONE 8:e69879. 2013
    ..In mice, systemically delivered CELiD DNA resulted in long-term, stable transgene expression in the liver. CELiD vectors represent a novel eukaryotic alternative to bacterial plasmid DNA. ..
  16. ncbi request reprint Insect cells as a factory to produce adeno-associated virus type 2 vectors
    Masashi Urabe
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
    Hum Gene Ther 13:1935-43. 2002
    ..This robust system provides a simple, cost-effective method for AAV vector production...
  17. ncbi request reprint Profiles of PrKX expression in developmental mouse embryo and human tissues
    Wei Li
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
    J Histochem Cytochem 53:1003-9. 2005
    ..Very low protein levels were found in heart tissues from a 5-month-old fetus and from an adult, whereas PrKX proteins were more abundant in fetal brain, kidney, and liver tissues compared with adult samples of the same tissue type...
  18. pmc Biochemical characterization of Junonia coenia densovirus nonstructural protein NS-1
    Chuantian Ding
    Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892 1654, USA
    J Virol 76:338-45. 2002
    ....
  19. ncbi request reprint Recombinant adeno-associated virus serotype 2 vectors mediate stable interleukin 10 secretion from salivary glands into the bloodstream
    Seiichi Yamano
    Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA
    Hum Gene Ther 13:287-98. 2002
    ....
  20. ncbi request reprint Transposase-mediated construction of an integrated adeno-associated virus type 5 helper plasmid
    Richard H Smith
    National Heart, Lung, and Blood Institute, Bethesda, MD 20892 1654, USA
    Biotechniques 33:204-6, 208, 210-1. 2002
    ....
  21. ncbi request reprint The nuclease domain of adeno-associated virus rep coordinates replication initiation using two distinct DNA recognition interfaces
    Alison Burgess Hickman
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892 USA
    Mol Cell 13:403-14. 2004
    ..Use of these different interfaces generates the structural asymmetry necessary to regulate later events in viral replication and integration...
  22. pmc Scalable generation of high-titer recombinant adeno-associated virus type 5 in insect cells
    Masashi Urabe
    Division of Genetic Therapeutics, Jichi Medical School, 3311 1 Yakushiji, Minami Kawachi, Tochigi 329 0498, Japan
    J Virol 80:1874-85. 2006
    ..These results indicated that rAAV5 can be generated in insect cells, although the difference in incorporated virion DNA may induce different expression patterns of the transgene...
  23. ncbi request reprint Widespread dispersion of adeno-associated virus serotype 1 and adeno-associated virus serotype 6 vectors in the rat central nervous system and in human glioblastoma multiforme xenografts
    Peter C Huszthy
    Haukeland University Hospital, N 5021 Bergen, Norway
    Hum Gene Ther 16:381-92. 2005
    ..The presented results suggest a potential therapeutic role for AAV1 and AAV6 vectors in gene therapy for GBM and also for other CNS malignancies...
  24. ncbi request reprint Recombinant adeno-associated virus serotypes 2- and 5-mediated gene transfer in the mammalian brain: quantitative analysis of heparin co-infusion
    Mihail Y Mastakov
    Functional Genomics and Translational Neuroscience Laboratory, Division of Molecular Medicine, Faculty of Medical and Health Sciences, University of Auckland, New Zealand
    Mol Ther 5:371-80. 2002
    ....
  25. pmc Immunological aspects of recombinant adeno-associated virus delivery to the mammalian brain
    Mihail Y Mastakov
    Functional Genomics and Translational Neuroscience Laboratory, Division of Molecular Medicine, Faculty of Medical and Health Sciences, University of Auckland, New Zealand
    J Virol 76:8446-54. 2002
    ..This study provides guidelines for more rational design of gene transfer studies in the rodent brain and, in addition, suggests the use of repeated administration of rAAV as a viable form of therapy for the treatment of chronic diseases...
  26. ncbi request reprint Adeno-associated virus type 5: transduction efficiency and cell-type specificity in the primate retina
    Andrew J Lotery
    Department of Ophthalmology, Roy J and Lucille A Carver College of Medicine, Iowa City, IA 52242, USA
    Hum Gene Ther 14:1663-71. 2003
    ..Our results support the utility of AAV5 for rod photoreceptor degeneration therapies...
  27. ncbi request reprint Recombinant adeno-associated virus vector for gene transfer to the transplanted rat heart
    Johannes M Schirmer
    William J von Liebig Transplant Center, Mayo Clinic, Rochester, MN 55905, USA
    Transpl Int 20:550-7. 2007
    ..This study demonstrates cardiac rAAV gene transduction with a cold perfusion preservation system of the donor heart. These data show that AAV-1 is superior to AAV-2 and AAV-5 for this purpose and that durable expression is achievable...
  28. ncbi request reprint RNAi suppresses polyglutamine-induced neurodegeneration in a model of spinocerebellar ataxia
    Haibin Xia
    Program in Gene Therapy, University of Iowa, Iowa City, Iowa, USA
    Nat Med 10:816-20. 2004
    ..Our data demonstrate in vivo the potential use of RNAi as therapy for dominant neurodegenerative disease...
  29. doi request reprint Glucose- and metabolically regulated hepatic insulin gene therapy for diabetes
    Paul Yueh Jen Hsu
    School of Pharmacy, College of Medicine, National Taiwan University, Taipei, 100, Taiwan
    Pharm Res 25:1460-8. 2008
    ..The purpose of this study was to examine glucose- and metabolically modulation of insulin secretion by rAAV-mediated gene delivery in vitro and in vivo...
  30. ncbi request reprint Adeno-associated virus (AAV) serotypes 2, 4 and 5 display similar transduction profiles and penetrate solid tumor tissue in models of human glioma
    Frits Thorsen
    Department of Biomedicine, Section of Anatomy and Cell Biology, University of Bergen, and Department of Oncology and Medical Physics, Haukeland University Hospital, Bergen, Norway
    J Gene Med 8:1131-40. 2006
    ..In the present work, the transduction efficacy of AAV serotypes 4 and 5 were compared to AAV2, both in vitro and in intracranial GBM xenografts derived from patient biopsies implanted into nude rats...