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Overview of Current Scalable Methods for Purification of Viral Vectors

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Viral Vectors for Gene Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 737))

Abstract

As a result of the growing interest in the use of viruses for gene therapy and vaccines, many virus-based products are being developed. The manufacturing of viruses poses new challenges for process developers and regulating authorities that need to be addressed to ensure quality, efficacy, and safety of the final product. The design of suitable purification strategies will depend on a multitude of variables including the vector production system and the nature of the virus. In this chapter, we provide an overview of the most commonly used purification methods for viral gene therapy vectors. Current chromatography options available for large-scale purification of γ-retrovirus, lentivirus, adenovirus, adeno-associated virus, herpes simplex virus, baculovirus, and poxvirus vectors are presented.

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References

  1. Altaras, N. E., Aunins, J. G., Evans, R. K., Kamen, A., Konz, J. O., and Wolf, J. J. (2005) Production and formulation of adenovirus vectors, Adv Biochem Eng Biotechnol 99, 193–260.

    PubMed  CAS  Google Scholar 

  2. Burova, E., and Ioffe, E. (2005) Chromatographic purification of recombinant adenoviral and adeno-associated viral vectors: methods and implications, Gene Ther 12 Suppl 1, S5–17.

    Google Scholar 

  3. Lusky, M. (2005) Good manufacturing practice production of adenoviral vectors for clinical trials, Hum Gene Ther 16, 281–291.

    Article  PubMed  CAS  Google Scholar 

  4. Segura, M. M., Alba, R., Bosch, A., and Chillon, M. (2008) Advances in helper-dependent adenoviral vector research, Curr Gene Ther 8, 222–235.

    Article  PubMed  CAS  Google Scholar 

  5. Rodrigues, T., Carrondo, M. J., Alves, P. M., and Cruz, P. E. (2007) Purification of retroviral vectors for clinical application: biological implications and technological challenges, J Biotechnol 127, 520–541.

    Article  PubMed  CAS  Google Scholar 

  6. Segura, M. M., Kamen, A., and Garnier, A. (2006) Downstream processing of oncoretroviral and lentiviral gene therapy vectors, Biotechnol Adv 24, 321–337.

    Article  PubMed  Google Scholar 

  7. Green, A. P., Huang, J. J., Scott, M. O., Kierstead, T. D., Beaupre, I., Gao, G. P., and Wilson, J. M. (2002) A new scalable method for the purification of recombinant adenovirus vectors, Hum Gene Ther 13, 1921–1934.

    Article  PubMed  CAS  Google Scholar 

  8. Lyddiatt, A., and O’Sullivan, D. A. (1998) Biochemical recovery and purification of gene therapy vectors, Curr Opin Biotechnol 9, 177–185.

    Article  PubMed  CAS  Google Scholar 

  9. Reeves, L., and Cornetta, K. (2000) Clinical retroviral vector production: step filtration using clinically approved filters improves titers, Gene Ther 7, 1993–1998.

    Article  PubMed  CAS  Google Scholar 

  10. Le Doux, J. M., Morgan, J. R., Snow, R. G., and Yarmush, M. L. (1996) Proteoglycans secreted by packaging cell lines inhibit retrovirus infection, J Virol 70, 6468–6473.

    PubMed  Google Scholar 

  11. Burns, J. C., Friedmann, T., Driever, W., Burrascano, M., and Yee, J. K. (1993) Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells, Proc Natl Acad Sci USA 90, 8033–8037.

    Article  PubMed  CAS  Google Scholar 

  12. Pham, L., Ye, H., Cosset, F. L., Russell, S. J., and Peng, K. W. (2001) Concentration of viral vectors by co-precipitation with calcium phosphate, J Gene Med 3, 188–194.

    Article  PubMed  CAS  Google Scholar 

  13. Schagen, F. H., Rademaker, H. J., Rabelink, M. J., van Ormondt, H., Fallaux, F. J., van der Eb, A. J., and Hoeben, R. C. (2000) Ammonium sulphate precipitation of recombinant adenovirus from culture medium: an easy method to increase the total virus yield, Gene Ther 7, 1570–1574.

    Article  PubMed  CAS  Google Scholar 

  14. Pedro, L., Soares, S. S., and Ferreira, G. N. M. (2008) Purification of bionanoparticles, Chem. Eng. Technol. 31, 815–825.

    Article  CAS  Google Scholar 

  15. Zhang, B., Xia, H. Q., Cleghorn, G., Gobe, G., West, M., and Wei, M. Q. (2001) A highly efficient and consistent method for harvesting large volumes of high-titre lentiviral vectors, Gene Ther 8, 1745–1751.

    Article  PubMed  CAS  Google Scholar 

  16. Kamen, A., and Henry, O. (2004) Development and optimization of an adenovirus production process, J Gene Med 6 Suppl 1, S184–192.

    Google Scholar 

  17. Koolman, J., and Rohm, K. H. (2004) “Organelles”. In Color Atlas of Biochemistry, 2nd ed., pp 196–234, Thieme Medical Publishers, Stuttgart-New York.

    Google Scholar 

  18. Round, J. J., Liptak, R. A., and McGregor, W. C. (1981) Continuous-flow ultracentrifugation in preparative biochemistry, Ann N Y Acad Sci 369, 265–274.

    Article  PubMed  CAS  Google Scholar 

  19. Reimer, C. B., Baker, R. S., Van Frank, R. M., Newlin, T. E., Cline, G. B., and Anderson, N. G. (1967) Purification of large quantities of influenza virus by density gradient centrifugation, J Virol 1, 1207–1216.

    PubMed  CAS  Google Scholar 

  20. Huyghe, B. G., Liu, X., Sutjipto, S., Sugarman, B. J., Horn, M. T., Shepard, H. M., Scandella, C. J., and Shabram, P. (1995) Purification of a type 5 recombinant adenovirus encoding human p53 by column chromatography, Hum Gene Ther 6, 1403–1416.

    Article  PubMed  CAS  Google Scholar 

  21. Blanche, F., Cameron, B., Barbot, A., Ferrero, L., Guillemin, T., Guyot, S., Somarriba, S., and Bisch, D. (2000) An improved anion-exchange HPLC method for the detection and purification of adenoviral particles, Gene Ther 7, 1055–1062.

    Article  PubMed  CAS  Google Scholar 

  22. Peixoto, C., Ferreira, T. B., Carrondo, M. J., Cruz, P. E., and Alves, P. M. (2006) Purification of adenoviral vectors using expanded bed chromatography, J Virol Methods 132, 121–126.

    Article  PubMed  CAS  Google Scholar 

  23. Vellekamp, G., Porter, F. W., Sutjipto, S., Cutler, C., Bondoc, L., Liu, Y. H., Wylie, D., Cannon-Carlson, S., Tang, J. T., Frei, A., Voloch, M., and Zhuang, S. (2001) Empty capsids in column-purified recombinant adenovirus preparations, Hum Gene Ther 12, 1923–1936.

    Article  PubMed  CAS  Google Scholar 

  24. Konz, J. O., Lee, A. L., Lewis, J. A., and Sagar, S. L. (2005) Development of a purification process for adenovirus: controlling virus aggregation to improve the clearance of host cell DNA, Biotechnol Prog 21, 466–472.

    Article  PubMed  CAS  Google Scholar 

  25. Peixoto, C., Ferreira, T. B., Sousa, M. F., Carrondo, M. J., and Alves, P. M. (2008) Towards purification of adenoviral vectors based on membrane technology, Biotechnol Prog 24, 1290–1296.

    Article  PubMed  CAS  Google Scholar 

  26. Urabe, M., Xin, K. Q., Obara, Y., Nakakura, T., Mizukami, H., Kume, A., Okuda, K., and Ozawa, K. (2006) Removal of empty capsids from type 1 adeno-associated virus vector stocks by anion-exchange chromatography potentiates transgene expression, Mol Ther 13, 823–828.

    Article  PubMed  CAS  Google Scholar 

  27. Zolotukhin, S., Potter, M., Zolotukhin, I., Sakai, Y., Loiler, S., Fraites, T. J., Jr., Chiodo, V. A., Phillipsberg, T., Muzyczka, N., Hauswirth, W. W., Flotte, T. R., Byrne, B. J., and Snyder, R. O. (2002) Production and purification of serotype 1, 2, and 5 recombinant adeno-associated viral vectors, Methods 28, 158–167.

    Article  PubMed  CAS  Google Scholar 

  28. Qu, G., Bahr-Davidson, J., Prado, J., Tai, A., Cataniag, F., McDonnell, J., Zhou, J., Hauck, B., Luna, J., Sommer, J. M., Smith, P., Zhou, S., Colosi, P., High, K. A., Pierce, G. F., and Wright, J. F. (2007) Separation of adeno-associated virus type 2 empty particles from genome containing vectors by anion-exchange column chromatography, J Virol Methods 140, 183–192.

    Article  PubMed  CAS  Google Scholar 

  29. Kaludov, N., Handelman, B., and Chiorini, J. A. (2002) Scalable purification of adeno-associated virus type 2, 4, or 5 using ion-exchange chromatography, Hum Gene Ther 13, 1235–1243.

    Article  PubMed  CAS  Google Scholar 

  30. Gao, G., Qu, G., Burnham, M. S., Huang, J., Chirmule, N., Joshi, B., Yu, Q. C., Marsh, J. A., Conceicao, C. M., and Wilson, J. M. (2000) Purification of recombinant adeno-associated virus vectors by column chromatography and its performance in vivo, Hum Gene Ther 11, 2079–2091.

    Article  PubMed  CAS  Google Scholar 

  31. O’Riordan, C. R., Lachapelle, A. L., Vincent, K. A., and Wadsworth, S. C. (2000) Scaleable chromatographic purification process for recombinant adeno-associated virus (rAAV), J Gene Med 2, 444–454.

    Article  PubMed  Google Scholar 

  32. Brument, N., Morenweiser, R., Blouin, V., Toublanc, E., Raimbaud, I., Cherel, Y., Folliot, S., Gaden, F., Boulanger, P., Kroner-Lux, G., Moullier, P., Rolling, F., and Salvetti, A. (2002) A versatile and scalable two-step ion-exchange chromatography process for the purification of recombinant adeno-associated virus serotypes-2 and -5, Mol Ther 6, 678–686.

    Article  PubMed  CAS  Google Scholar 

  33. Davidoff, A. M., Ng, C. Y., Sleep, S., Gray, J., Azam, S., Zhao, Y., McIntosh, J. H., Karimipoor, M., and Nathwani, A. C. (2004) Purification of recombinant adeno-associated virus type 8 vectors by ion exchange ­chromatography generates clinical grade ­vector stock, J Virol Methods 121, 209–215.

    Article  PubMed  CAS  Google Scholar 

  34. Smith, R. H., Ding, C., and Kotin, R. M. (2003) Serum-free production and column purification of adeno-associated virus type 5, J Virol Methods 114, 115–124.

    Article  PubMed  CAS  Google Scholar 

  35. Vicente, T., Peixoto, C., Carrondo, M. J., and Alves, P. M. (2009) Purification of recombinant baculoviruses for gene therapy using membrane processes, Gene Ther 16, 766–775.

    Article  PubMed  CAS  Google Scholar 

  36. Scherr, M., Battmer, K., Eder, M., Schule, S., Hohenberg, H., Ganser, A., Grez, M., and Blomer, U. (2002) Efficient gene transfer into the CNS by lentiviral vectors purified by anion exchange chromatography, Gene Ther 9, 1708–1714.

    Article  PubMed  CAS  Google Scholar 

  37. Yamada, K., McCarty, D. M., Madden, V. J., and Walsh, C. E. (2003) Lentivirus vector purification using anion exchange HPLC leads to improved gene transfer, Biotechniques 34, 1074–1078, 1080.

    Google Scholar 

  38. Slepushkin, V., Chang, N., Cohen, R., Gan, Y., Jiang, B., Deausen, E., Berlinger, D., Binder, G., Andre, K., Humeau, L., and Dropulic, B. (2003) Large-scale purification of a lentiviral vector by size exclusion chromatography or mustang Q ion exchange capsule, Bioprocessing J. 2, 89–95.

    Google Scholar 

  39. Kutner, R. H., Puthli, S., Marino, M. P., and Reiser, J. (2009) Simplified production and concentration of HIV-1-based lentiviral vectors using HYPERFlask vessels and anion exchange membrane chromatography, BMC Biotechnol 9, 10.

    Article  PubMed  Google Scholar 

  40. Rodrigues, T., Carvalho, A., Carmo, M., Carrondo, M. J., Alves, P. M., and Cruz, P. E. (2007) Scaleable purification process for gene therapy retroviral vectors, J Gene Med 9, 233–243.

    Article  PubMed  CAS  Google Scholar 

  41. Rodrigues, T., Carvalho, A., Roldao, A., Carrondo, M. J., Alves, P. M., and Cruz, P. E. (2006) Screening anion-exchange chromatographic matrices for isolation of onco-retroviral vectors, J Chromatogr B Analyt Technol Biomed Life Sci 837, 59–68.

    Article  PubMed  CAS  Google Scholar 

  42. Zolotukhin, S., Byrne, B. J., Mason, E., Zolotukhin, I., Potter, M., Chesnut, K., Summerford, C., Samulski, R. J., and Muzyczka, N. (1999) Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield, Gene Ther 6, 973–985.

    Article  PubMed  CAS  Google Scholar 

  43. Chahal, P. S., Aucoin, M. G., and Kamen, A. (2007) Primary recovery and chromatographic purification of adeno-associated virus type 2 produced by baculovirus/insect cell system, J Virol Methods 139, 61–70.

    Article  PubMed  CAS  Google Scholar 

  44. Wu, C., Soh, K. Y., and Wang, S. (2007) ­Ion-exchange membrane chromatography method for rapid and efficient purification of recombinant baculovirus and baculovirus gp64 protein, Hum Gene Ther 18, 665–672.

    Article  PubMed  CAS  Google Scholar 

  45. Kuiper, M., Sanches, R. M., Walford, J. A., and Slater, N. K. (2002) Purification of a functional gene therapy vector derived from Moloney murine leukaemia virus using membrane filtration and ceramic hydroxyapatite chromatography, Biotechnol Bioeng 80, 445–453.

    Article  PubMed  CAS  Google Scholar 

  46. Smith, R. H., Levy, J. R., and Kotin, R. M. (2009) A simplified baculovirus-AAV expression vector system coupled with one-step affinity purification yields high-titer rAAV stocks from insect cells, Mol Ther 17, 1888–1896.

    Article  PubMed  CAS  Google Scholar 

  47. Grimm, D., Kern, A., Rittner, K., and Kleinschmidt, J. A. (1998) Novel tools for production and purification of recombinant adenoassociated virus vectors, Hum Gene Ther 9, 2745–2760.

    Article  PubMed  CAS  Google Scholar 

  48. Clark, K. R., Liu, X., McGrath, J. P., and Johnson, P. R. (1999) Highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses, Hum Gene Ther 10, 1031–1039.

    Article  PubMed  CAS  Google Scholar 

  49. Anderson, R., Macdonald, I., Corbett, T., Whiteway, A., and Prentice, H. G. (2000) A method for the preparation of highly purified adeno-associated virus using affinity column chromatography, protease digestion and solvent extraction, J Virol Methods 85, 23–34.

    Article  PubMed  CAS  Google Scholar 

  50. Harris, J. D., Beattie, S. G., and Dickson, J. G. (2003) Novel tools for production and purification of recombinant adeno-associated viral vectors, Methods Mol Med 76, 255–267.

    PubMed  CAS  Google Scholar 

  51. Segura, M. M., Kamen, A., Trudel, P., and Garnier, A. (2005) A novel purification strategy for retrovirus gene therapy vectors using heparin affinity chromatography, Biotechnol Bioeng 90, 391–404.

    Article  PubMed  Google Scholar 

  52. Segura, M. M., Garnier, A., Durocher, Y., Coelho, H., and Kamen, A. (2007) Production of lentiviral vectors by large-scale transient transfection of suspension cultures and affinity chromatography purification, Biotechnol Bioeng 98, 789–799.

    Article  PubMed  CAS  Google Scholar 

  53. Wolff, M. W., Venzke, C., Zimmermann, A., Post Hansen, S., Djurup, R., Faber, R., and Reichl, U. (2007) Affinity Chromatography Of Cell Culture Derived Vaccinia Virus, in American Institute of Chemical Engineers, Salt Lake City

    Google Scholar 

  54. Wolff, M. W., Sievers, C., Lehmann, S., Opitz, L., Post Hansen, S., Djurup, R., Faber, R., and Reichl, U. (2008) Capturing of Cell Culture Derived Vaccinia Virus by Membrane Adsorbers, in American Institute of Chemical Engineers, Philadelphia.

    Google Scholar 

  55. Wolff, M. W., Sievers, C., Lehmann, S., Post Hansen, S., Faber, R., and Reichl, U. (2009) Cellufine® sulfate and heparin affinity chromatography to capture cell culture derived Vaccinia Virus particles, in 21st Meeting of the European Society for Animal Cell Technology.

    Google Scholar 

  56. Auricchio, A., O’Connor, E., Hildinger, M., and Wilson, J. M. (2001) A single-step affinity column for purification of serotype-5 based adeno-associated viral vectors, Mol Ther 4, 372–374.

    Article  PubMed  CAS  Google Scholar 

  57. Koerber, J. T., Jang, J. H., Yu, J. H., Kane, R. S., and Schaffer, D. V. (2007) Engineering adeno-associated virus for one-step purification via immobilized metal affinity chromatography, Hum Gene Ther 18, 367–378.

    Article  PubMed  CAS  Google Scholar 

  58. Hu, Y., Tsai, C., Chung, Y., Lu, J., and Hsu, J. T. (2003) Generation of chimeric baculovirus with histidine-tags displayed on the envelope and its purification using immobilized metal affinity chromatography, Enzyme and Microbial Technology 33, 445–452.

    Article  CAS  Google Scholar 

  59. Jiang, C., Glorioso, J. C., and Ataai, M. (2006) Presence of imidazole in loading buffer prevents formation of free radical in immobilized metal affinity chromatography and dramatically improves the recovery of herpes simplex virus type 1 gene therapy vectors, J Chromatogr A 1121, 40–45.

    Article  PubMed  CAS  Google Scholar 

  60. Jiang, C., Wechuck, J. B., Goins, W. F., Krisky, D. M., Wolfe, D., Ataai, M. M., and Glorioso, J. C. (2004) Immobilized cobalt affinity chromatography provides a novel, efficient method for herpes simplex virus type 1 gene vector purification, J Virol 78, 8994–9006.

    Article  PubMed  CAS  Google Scholar 

  61. Ye, K., Jin, S., Ataai, M. M., Schultz, J. S., and Ibeh, J. (2004) Tagging retrovirus vectors with a metal binding peptide and one-step purification by immobilized metal affinity chromatography, J Virol 78, 9820–9827.

    Article  PubMed  CAS  Google Scholar 

  62. Yu, J. H., and Schaffer, D. V. (2006) Selection of novel vesicular stomatitis virus glycoprotein variants from a peptide insertion library for enhanced purification of retroviral and lentiviral vectors, J Virol 80, 3285–3292.

    Article  PubMed  CAS  Google Scholar 

  63. Cheeks, M. C., Kamal, N., Sorrell, A., Darling, D., Farzaneh, F., and Slater, N. K. (2009) Immobilized metal affinity chromatography of histidine-tagged lentiviral vectors using monolithic adsorbents, J Chromatogr A 1216, 2705–2711.

    Article  PubMed  CAS  Google Scholar 

  64. Stachler, M. D., and Bartlett, J. S. (2006) Mosaic vectors comprised of modified AAV1 capsid proteins for efficient vector purification and targeting to vascular endothelial cells, Gene Ther 13, 926–931.

    PubMed  CAS  Google Scholar 

  65. Parrott, M. B., Adams, K. E., Mercier, G. T., Mok, H., Campos, S. K., and Barry, M. A. (2003) Metabolically biotinylated adenovirus for cell targeting, ligand screening, and vector purification, Mol Ther 8, 688–700.

    Article  PubMed  CAS  Google Scholar 

  66. Williams, S. L., Nesbeth, D., Darling, D. C., Farzaneh, F., and Slater, N. K. (2005) Affinity recovery of Moloney Murine Leukaemia Virus, J Chromatogr B Analyt Technol Biomed Life Sci 820, 111–119.

    Article  PubMed  CAS  Google Scholar 

  67. Williams, S. L., Eccleston, M. E., and Slater, N. K. (2005) Affinity capture of a biotinylated retrovirus on macroporous monolithic adsorbents: towards a rapid single-step purification process, Biotechnol Bioeng 89, 783–787.

    Article  PubMed  CAS  Google Scholar 

  68. Potter, M., Chesnut, K., Muzyczka, N., Flotte, T., and Zolotukhin, S. (2002) Streamlined large-scale production of recombinant adeno-associated virus (rAAV) vectors, Methods Enzymol 346, 413–430.

    Article  PubMed  CAS  Google Scholar 

  69. Transfiguracion, J., Jorio, H., Meghrous, J., Jacob, D., and Kamen, A. (2007) High yield purification of functional baculovirus vectors by size exclusion chromatography, J Virol Methods 142, 21–28.

    Article  PubMed  CAS  Google Scholar 

  70. Transfiguracion, J., Jaalouk, D. E., Ghani, K., Galipeau, J., and Kamen, A. (2003) Size-exclusion chromatography purification of high-titer vesicular stomatitis virus G glycoprotein-pseudotyped retrovectors for cell and gene therapy applications, Hum Gene Ther 14, 1139–1153.

    Article  PubMed  CAS  Google Scholar 

  71. Esser, M. T., Graham, D. R., Coren, L. V., Trubey, C. M., Bess, J. W., Jr., Arthur, L. O., Ott, D. E., and Lifson, J. D. (2001) Differential incorporation of CD45, CD80 (B7-1), CD86 (B7-2), and major histocompatibility complex class I and II molecules into human immunodeficiency virus type 1 virions and microvesicles: implications for viral pathogenesis and immune regulation, J Virol 75, 6173–6182.

    Article  PubMed  CAS  Google Scholar 

  72. Trubey, C. M., Chertova, E., Coren, L. V., Hilburn, J. M., Hixson, C. V., Nagashima, K., Lifson, J. D., and Ott, D. E. (2003) Quantitation of HLA class II protein incorporated into human immunodeficiency type 1 virions purified by anti-CD45 immunoaffinity depletion of microvesicles, J Virol 77, 12699–12709.

    Article  PubMed  CAS  Google Scholar 

  73. Andreadis, S. T., Roth, C. M., Le Doux, J. M., Morgan, J. R., and Yarmush, M. L. (1999) Large-scale processing of recombinant retroviruses for gene therapy, Biotechnol Prog 15, 1–11.

    Article  PubMed  CAS  Google Scholar 

  74. O’Keeffe, R. S., Johnston, M. D., and Slater, N. K. (1999) The affinity adsorptive recovery of an infectious herpes simplex virus vaccine, Biotechnol Bioeng 62, 537–545.

    Article  PubMed  Google Scholar 

  75. Stadler, J., Lemmens, R., and Nyhammar, T. (2004) Plasmid DNA purification, J Gene Med 6 Suppl 1, S54–66.

    Google Scholar 

  76. Kalbfuss, B., Wolff, M., Morenweiser, R., and Reichl, U. (2007) Purification of cell culture-derived human influenza A virus by size-exclusion and anion-exchange chromatography, Biotechnol Bioeng 96, 932–944.

    Article  PubMed  CAS  Google Scholar 

  77. Morenweiser, R. (2005) Downstream processing of viral vectors and vaccines, Gene Ther 12 Suppl 1, S103–110.

    Google Scholar 

  78. Ljunglof, A., Bergvall, P., Bhikhabhai, R., and Hjorth, R. (1999) Direct visualisation of plasmid DNA in individual chromatography adsorbent particles by confocal scanning laser microscopy, J Chromatogr A 844, 129–135.

    Article  PubMed  CAS  Google Scholar 

  79. Urthaler, J., Buchinger, W., and Necina, R. (2005) Improved downstream process for the production of plasmid DNA for gene therapy, Acta Biochim Pol 52, 703–711.

    PubMed  CAS  Google Scholar 

  80. Kaufmann, M. (1997) Unstable proteins: how to subject them to chromatographic separations for purification procedures, J Chromatogr B Biomed Sci Appl 699, 347–369.

    Article  PubMed  CAS  Google Scholar 

  81. McIntyre, M. (2001) Development of viral vectors for use in gene transfer trials; product characterization and quality concerns., Adenovirus Reference Material Working Group: http://www.wilbio.com.

  82. Okada, T., Nonaka-Sarukawa, M., Uchibori, R., Kinoshita, K., Hayashita-Kinoh, H., Nitahara-Kasahara, Y., Takeda, S., and Ozawa, K. (2009) Scalable purification of adeno-associated virus serotype 1 (AAV1) and AAV8 vectors, using dual ion-exchange adsorptive membranes, Hum Gene Ther 20, 1013–1021.

    Article  PubMed  CAS  Google Scholar 

  83. Palmer, D., and Ng, P. (2003) Improved system for helper-dependent adenoviral vector production, Mol Ther 8, 846–852.

    Article  PubMed  CAS  Google Scholar 

  84. Sakhuja, K., Reddy, P. S., Ganesh, S., Cantaniag, F., Pattison, S., Limbach, P., Kayda, D. B., Kadan, M. J., Kaleko, M., and Connelly, S. (2003) Optimization of the generation and propagation of gutless adenoviral vectors, Hum Gene Ther 14, 243–254.

    Article  PubMed  CAS  Google Scholar 

  85. Bess, J. W., Jr., Gorelick, R. J., Bosche, W. J., Henderson, L. E., and Arthur, L. O. (1997) Microvesicles are a source of contaminating cellular proteins found in purified HIV-1 preparations, Virology 230, 134–144.

    Article  PubMed  CAS  Google Scholar 

  86. Gluschankof, P., Mondor, I., Gelderblom, H. R., and Sattentau, Q. J. (1997) Cell membrane vesicles are a major contaminant of gradient-enriched human immunodeficiency virus type-1 preparations, Virology 230, 125–133.

    Article  PubMed  CAS  Google Scholar 

  87. Segura, M. M., Garnier, A., and Kamen, A. (2006) Purification and characterization of retrovirus vector particles by rate zonal ultracentrifugation, J Virol Methods 133, 82–91.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

The authors wish to thank Gavin Whissell for careful review of this manuscript. This work was supported by an NSERC Strategic Project grant and the NCE Canadian Stem Cell Network.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Center of Animal Biotechnology and Gene Therapy (CBATEG), Universitat Autònoma de Barcelona, Barcelona, Spain

    María Mercedes Segura

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  1. María Mercedes Segura
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  2. Amine A. Kamen
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  3. Alain Garnier
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Correspondence to María Mercedes Segura .

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Editors and Affiliations

  1. Généthon, rue de l'Internationale 1, Évry CX 2, 91002, France

    Otto-Wilhelm Merten

  2. Chemical & Bioprocess Engineering, University College Dublin, Belfield, Dublin, 4, Ireland

    Mohamed Al-Rubeai

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Segura, M.M., Kamen, A.A., Garnier, A. (2011). Overview of Current Scalable Methods for Purification of Viral Vectors. In: Merten, OW., Al-Rubeai, M. (eds) Viral Vectors for Gene Therapy. Methods in Molecular Biology, vol 737. Humana Press. https://doi.org/10.1007/978-1-61779-095-9_4

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  • DOI: https://doi.org/10.1007/978-1-61779-095-9_4

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