Manufacturing of recombinant adeno-associated viruses using mammalian expression platforms

Biotechnology Journal

Volumn 12, Issue 3, 2017, Pages

  Robert, Marc André ^a,b   Chahal, Parminder S ^b   Audy, Alexandre ^a,b   Kamen, Amine ^c   Gilbert, Rénald ^b   Gaillet, Bruno ^a  

^a UNIVERSITÉ LAVAL   (Canada)

^b NATIONAL RESEARCH COUNCIL CANADA   (Canada)

^c MCGILL UNIVERSITY   (Canada)

Author keywords

Gene therapy; HEK293; Large scale manufacturing; Mammalian cells; Recombinant adeno associated viruses

Indexed keywords

CELLS; CYTOLOGY; GENE THERAPY; GENES; MAMMALS; MANUFACTURE; PURIFICATION; VIRUSES;

HEK-293; LARGE-SCALE MANUFACTURING; MAMMALIAN CELLS; MANUFACTURING PRACTICES; RECOMBINANT ADENO-ASSOCIATED VIRUS; SMALL-SCALE PRODUCTION; TANGENTIAL FLOW FILTRATIONS; TRANSIENT TRANSFECTION;

MOLECULAR BIOLOGY;

ADENO ASSOCIATED VIRUS VECTOR; CESIUM CHLORIDE; IODIXANOL;

ADENO ASSOCIATED VIRUS; ANALYTIC METHOD; CELL LYSATE; CHROMATOGRAPHY; GENETIC TRANSFECTION; HEK293 CELL LINE; HERPES SIMPLEX VIRUS; MAMMAL CELL; MICROFILTRATION; NONHUMAN; PRIORITY JOURNAL; REVIEW; ULTRACENTRIFUGATION; VIRAL GENE THERAPY; VIRUS EXPRESSION; VIRUS RECOMBINANT; BIOSYNTHESIS; DEPENDOPARVOVIRUS; GENE THERAPY; GENE VECTOR; GENETICS; HUMAN; ISOLATION AND PURIFICATION; PROCEDURES; VIRUS CULTURE;

DEPENDOVIRUS; GENETIC THERAPY; GENETIC VECTORS; HEK293 CELLS; HUMANS; TRANSFECTION; VIRUS CULTIVATION;

EID: 85012070511     PISSN: 18606768     EISSN: 18607314     Source Type: Journal    
DOI: 10.1002/biot.201600193     Document Type: Review

References (138)

1. Wagner, J. A., Reynolds, T., Moran, M. L., Moss, R. B. et al., Efficient and persistent gene transfer of AAV-CFTR in maxillary sinus. Lancet 1998, 351, 1702–1703.
2. Wagner, J. A., Messner, A. H., Moran, M. L., Daifuku, R. et al., Safety and biological efficacy of an adeno-associated virus vector-cystic fibrosis transmembrane regulator (AAV-CFTR) in the cystic fibrosis maxillary sinus. Laryngoscope 1999, 109, 266–274.
3. Stone, D., Liu, Y., Li, Z.-Y., Strauss, R. et al., Biodistribution and safety profile of recombinant adeno-associated virus serotype 6 vectors following intravenous delivery. J. Virol. 2008, 82, 7711–7715.
4. Sun, J. Y., Anand-Jawa, V., Chatterjee, S., Wong, K. K., Immune responses to adeno-associated virus and its recombinant vectors. Gene Ther. 2003, 10, 964–976.
5. Zincarelli, C., Soltys, S., Rengo, G., Rabinowitz, J. E., Analysis of AAV serotypes 1–9 mediated gene expression and tropism in mice after systemic injection. Mol. Ther. 2008, 16, 1073–1080.
6. Jacobson, S. G., Cideciyan, A. V, Ratnakaram, R., Heon, E. et al., Gene therapy for Leber Congenital Amaurosis caused by RPE65 mutations: Safety and efficacy in fifteen children and adults followed up to three years. Arch. Ophthalmol. 2012, 130, 9–24.
7. Ferreira, V., Petry, H., Salmon, F., Immune responses to AAV-vectors, The Glybera example from bench to bedside. Front. Immunol. 2014, 5, 1–15.
8. Cecchini, S., Virag, T., Kotin, R. M., Reproducible high yields of recombinant adeno-associated virus produced using invertebrate cells in 0.02- to 200-liter cultures. Hum. Gene Ther. 2011, 22, 1021–1030.
9. Elias, C. B., Zeiser, A., Bédard, C., Kamen, A. A., Enhanced growth of Sf-9 cells to a maximum density of 5.2 × 107 cells per mL and production of β-galactosidase at high cell density by fed batch culture. Biotechnol. Bioeng. 2000, 68, 381–388.
10. Aucoin, M. G., Perrier, M., Kamen, A. A., Critical assessment of current adeno-associated viral vector production and quantification methods. Biotechnol. Adv. 2008, 26, 73–88.
11. Merten, O. W., Gaillet, B., Viral vectors for gene therapy and gene modification approaches. Biochem. Eng. J. 2016, 108, 98–115.
12. Clément, N., Grieger, J. C., Manufacturing of recombinant adeno-associated viral vectors for clinical trials. Mol. Ther. Methods Clin. Dev. 2016, 3, 16002.
13. Thomas, D. L., Wang, L., Niamke, J., Liu, J. et al., Scalable recombinant adeno-associated virus production using recombinant herpes simplex virus type 1 coinfection of suspension-adapted mammalian cells. Hum. Gene Ther. 2009, 20, 861–870.
14. Grieger, J. C., Soltys, S. M., Samulski, R. J., Production of recombinant adeno-associated virus vectors using suspension HEK293 cells and continuous harvest of vector from the culture media for GMP FIX and FLT1 clinical vector. Mol. Ther. 2016, 24, 287–297.
15. Thorne, B. A., Takeya, R. K., Peluso, R. W., Manufacturing recombinant adeno-associated viral vectors from producer cell clones. Hum. Gene Ther. 2009, 20, 707–714.
16. Balakrishnan, B., Jayandharan, G. R., Basic biology of adeno-associated virus (AAV) vectors used in gene therapy. Curr. Gene Ther. 2014, 14, 86–100.
17. Weitzman, M. D., Linden, R. M., Adeno-associated virus biology. Methods Mol. Biol. 2011, 807, 1–23.
18. Hölscher, C., Kleinschmidt, J. A., Bürkle, A., High-level expression of adeno-associated virus (AAV) Rep78 or Rep68 protein is sufficient for infectious-particle formation by a rep-negative AAV mutant. J. Virol. 1995, 69, 6880–6885.
19. Hamilton, H., Gomos, J., Berns, K. I., Falck-Pedersen, E., Adeno-associated virus site-specific integration and AAVS1 disruption. J. Virol. 2004, 78, 7874–7882.
20. Dong, B., Nakai, H., Xiao, W., Characterization of genome integrity for oversized recombinant AAV vector. Mol. Ther. 2010, 18, 87–92.
21. Wu, Z., Yang, H., Colosi, P., Effect of genome size on AAV vector packaging. Mol. Ther. 2010, 18, 80–86.
22. Brister, J. R., Muzyczka, N., Rep-mediated nicking of the adeno-associated virus origin requires two biochemical activities, DNA helicase activity and transesterification. J. Virol. 1999, 73, 9325–9336.
23. Wang, Z., Ma, H.-I., Li, J., Sun, L. et al., Rapid and highly efficient transduction by double-stranded adeno-associated virus vectors in vitro and in vivo. Gene Ther. 2003, 10, 2105–2111.
24. McCarty, D. M., Self-complementary AAV vectors; Advances and applications. Mol. Ther. 2008, 16, 1648–1656.
25. Summerford, C., Samulski, R. J., Membrane-associated heparan sulfate proteoglycan is a receptor for adeno-associated virus type 2 virions. J. Virol. 1998, 72, 1438–1445.
26. Croyle, M. A., Cheng, X., Wilson, J. M., Development of formulations that enhance physical stability of viral vectors for gene therapy. Gene Ther. 2001, 8, 1281–1290.
27. Gruntman, A. M., Su, L., Su, Q., Gao, G. et al., Stability and compatibility of recombinant adeno-associated virus under conditions commonly encountered in human gene therapy trials. Hum. Gene Ther. Methods 2015, 26, 71–76.
28. Potter, M., Lins, B., Mietzsch, M., Heilbronn, R. et al., A simplified purification protocol for recombinant adeno-associated virus vectors. Mol. Ther. Methods Clin. Dev. 2014, 1, 14034.
29. Rayaprolu, V., Kruse, S., Kant, R., Venkatakrishnan, B. et al., Comparative analysis of adeno associated virus capsid stability and dynamics. J. Virol. 2013, 87, 13150–13160.
30. Qiao, C., Li, J., Skold, A., Zhang, X., Xiao, X., Feasibility of generating adeno-associated virus packaging cell lines containing inducible adenovirus helper genes. J. Virol. 2002, 76, 1904–1913.
31. Gao, G., Lu, F., Sanmiguel, J. C., Tran, P. T. et al., Rep/Cap gene amplification and high-yield production of AAV in an A549 cell line expressing Rep/Cap. Mol. Ther. 2002, 5, 644–649.
32. Clément, N., Knop, D. R., Byrne, B. J., Large-scale adeno-associated viral vector production using a herpesvirus-based system enables manufacturing for clinical studies. Hum. Gene Ther. 2009, 20, 796–806.
33. Ayuso, E., Manufacturing of recombinant adeno-associated viral vectors: New technologies are welcome. Mol. Ther. Methods Clin. Dev. 2016, 3, 15049.
34. Merten, O.-W., Gény-Fiamma, C., Douar, a M., Current issues in adeno-associated viral vector production. Gene Ther. 2005, 12, 51–61.
35. Wright, J. F., Transient transfection methods for clinical adeno-associated viral vector production. Hum. Gene Ther. 2009, 20, 698–706.
36. Hill, M., Iaa, N. W., Graham, F., Smiley, R. et al., Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J. Gen. Virol. 1977, 36, 59–74.
37. Xiao, X., Li, J., Samulski, R. J., Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus. J. Virol. 1998, 72, 2224–2232.
38. Grimm, D., Kern, A., Rittner, K., Kleinschmidt, J. A., Novel tools for production and purification of recombinant adenoassociated virus vectors. Hum. Gene Ther. 1998, 9, 2745–2760.
39. Collaco, R. F., Cao, X., Trempe, J. P., A helper virus-free packaging system for recombinant adeno-associated virus vectors. Gene 1999, 238, 397–405.
40. Emmerling, V. V., Pegel, A., Milian, E. G., Venereo-Sanchez, A. et al., Rational plasmid design and bioprocess optimization to enhance recombinant adeno-associated virus (AAV) productivity in mammalian cells. Biotechnol. J. 2016, 11, 290–297.
41. Ling, C., Yin, Z., Li, J., Zhang, D. et al., Strategies to generate high-titer, high-potency recombinant AAV3 serotype vectors. Mol. Ther. Methods Clin. Dev. 2016, 3, 16029.
42. van der Loo, J. C. M., Wright, J. F., Progress and challenges in viral vector manufacturing. Hum. Mol. Genet. 2015, 2, 1–11.
43. Lock, M., Alvira, M., Vandenberghe, L. H., Samanta, A. et al., Rapid, simple, and versatile manufacturing of recombinant adeno-associated viral vectors at scale. Hum. Gene Ther. 2010, 21, 1259–1271.
44. Rashnonejad, A., Chermahini, G. A., Li, S., Ozkinay, F., Gao, G., Large-scale production of adeno-associated viral vector serotype-9 carrying the human survival motor neuron gene. Mol. Biotechnol. 2016, 58, 30–36.
45. Durocher, Y., Pham, P. L., St-Laurent, G., Jacob, D. et al., Scalable serum-free production of recombinant adeno-associated virus type 2 by transfection of 293 suspension cells. J. Virol. Methods 2007, 144, 32–40.
46. Hildinger, M., Baldi, L., Stettler, M., Wurm, F. M., High-titer, serum-free production of adeno-associated virus vectors by polyethyleneimine-mediated plasmid transfection in mammalian suspension cells. Biotechnol. Lett. 2007, 29, 1713–1721.
47. Chahal, P. S., Schulze, E., Tran, R., Montes, J., Kamen, A. a., Production of adeno-associated virus (AAV) serotypes by transient transfection of HEK293 cell suspension cultures for gene delivery. J. Virol. Methods 2014, 196, 163–173.
48. Jenny, C., Toublanc, E., Danos, O., Merten, O. W., Evaluation of a serum-free medium for the production of rAAV-2 using HeLa derived producer cells. Cytotechnology 2005, 49, 11–23.
49. Clark, K. R., Voulgaropoulou, F., Fraley, D. M., Johnson, P. R., Cell lines for the production of recombinant adeno-associated virus. Hum. Gene Ther. 1995, 6, 1329–1341.
50. Farson, D., Harding, T. C., Tao, L., Liu, J. et al., Development and characterization of a cell line for large-scale, serum-free production of recombinant adeno-associated viral vectors. J. Gene Med. 2004, 6, 1369–1381.
51. Martin, J., Frederick, A., Luo, Y., Jackson, R. et al., Generation and characterization of adeno-associated virus producer cell lines for research and preclinical vector production. Hum. Gene Ther. Methods 2013, 24, 253–269.
52. Qiao, C., Wang, B., Zhu, X., Li, J., Xiao, X., A novel gene expression control system and its use in stable, high-titer 293 cell-based adeno-associated virus packaging cell lines. J. Virol. 2002, 76, 13015–13027.
53. Yuan, Z., Qiao, C., Hu, P., Li, J., Xiao, X., A versatile adeno-associated virus vector producer cell line method for scalable vector production of different serotypes. Hum. Gene Ther. 2011, 22, 613–624.
54. Gao, G. P., Qu, G., Faust, L. Z., Engdahl, R. K. et al., High-titer adeno-associated viral vectors from a Rep/Cap cell line and hybrid shuttle virus. Hum. Gene Ther. 1998, 9, 2353–2362.
55. Gao, G., Rep/Cap gene amplification and high-yield production of AAV in an A549 cell line expressing Rep/Cap. Mol. Ther. 2002, 5, 644–649.
56. Thorne, B. A., Takeya, R. K., Peluso, R. W., Manufacturing recombinant adeno-associated viral vectors from producer cell clones. Hum. Gene Ther. 2009, 20, 707–714.
57. Adamson-Small, L., Potter, M., Falk, D. J., Cleaver, B. et al., A scalable method for the production of high-titer and high-quality adeno-associated type 9 vectors using the HSV platform. Mol. Ther. Methods Clin. Dev. 2016, 3, 16031.
58. Kang, W., Wang, L., Harrell, H., Liu, J. et al., An efficient rHSV-based complementation system for the production of multiple rAAV vector serotypes. Gene Ther. 2009, 16, 229–239.
59. Conway, J. E., Rhys, C. M., Zolotukhin, I., Zolotukhin, S. et al., High-titer recombinant adeno-associated virus production utilizing a recombinant herpes simplex virus type I vector expressing AAV-2 Rep and Cap. Gene Ther. 1999, 6, 986–993.
60. Vandenberghe, L. H., Xiao, R., Lock, M., Lin, J. et al., Efficient serotype-dependent release of functional vector into the culture medium during adeno-associated virus manufacturing. Hum. Gene Ther. 2010, 21, 1251–1257.
61. Croughan, W. S., Behbehani, A. M., Comparative study of inactivation of herpes simplex virus types 1 and 2 by commonly used antiseptic agents. J. Clin. Microbiol. 1988, 26, 213–215.
62. Strobel, B., Miller, F. D., Rist, W., Lamla, T., Comparative analysis of cesium chloride- and iodixanol-based purification of recombinant adeno-associated virus (AAV) vectors for preclinical applications. Hum. Gene Ther. Methods 2015, 112, 1–29.
63. Shin, J.-H., Yue, Y., Duan, D., Recombinant adeno-associated viral vector production and purification. Methods Mol. Biol. 2012, 798, 267–284.
64. Zolotukhin, S., Byrne, B., Mason, E., Zolotukhin, I. et al., Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Ther. 1999, 6, 973–985.
65. Doria, M., Ferrara, A., Auricchio, A., AAV2/8 vectors purified from culture medium with a simple and rapid protocol transduce murine liver, muscle, and retina efficiently. Hum. Gene Ther. Methods 2013, 24, 392–398.
66. Dias Florencio, G., Precigout, G., Beley, C., Buclez, P.-O. et al., Simple downstream process based on detergent treatment improves yield and in vivo transduction efficacy of adeno-associated virus vectors. Mol. Ther. Methods Clin. Dev. 2015, 2, 15024.
67. Powers, A. D., Piras, B. A., Clark, R. K., Lockey, T., Meagher, M. M., Development and optimization of AAV hFIX particles by transient transfection in an iCELLis(®); fixed bed bioreactor. Hum. Gene Ther. Methods 2016, 27, 112–121.
68. Chahal, P. S., Aucoin, M. G., Kamen, A., Primary recovery and chromatographic purification of adeno-associated virus type 2 produced by baculovirus/insect cell system. J. Virol. Methods 2007, 139, 61–70.
69. Wright, J. F., Le, T., Prado, J., Bahr-Davidson, J. et al., Identification of factors that contribute to recombinant AAV2 particle aggregation and methods to prevent its occurrence during vector purification and formulation. Mol. Ther. 2005, 12, 171–178.
70. Qu, W., Wang, M., Wu, Y., Xu, R., Scalable downstream strategies for purification of recombinant adeno-associated virus vectors in light of the properties. Curr. Pharm. Biotechnol. 2015, 16, 684–695.
71. Qu, G., Bahr-Davidson, J., Prado, J., Tai, A. et al., Separation of adeno-associated virus type 2 empty particles from genome containing vectors by anion-exchange column chromatography. J. Virol. Methods 2007, 140, 183–192.
72. Smith, R. H., Yang, L., Kotin, R. M., Chromatography-based purification of adeno-associated virus. Methods Mol. Biol. 2008, 434, 37–54.
73. Zolotukhin, S., Potter, M., Zolotukhin, I., Sakai, Y. et al., Production and purification of serotype 1, 2, and 5 recombinant adeno-associated viral vectors. Methods 2002, 28, 158–167.
74. Kaludov, N., Handelman, B., Chiorini, J. A., Scalable purification of adeno-associated virus type 2, 4, or 5 using ion-exchange chromatography. Hum. Gene Ther. 2002, 13, 1235–1243.
75. Brument, N., Morenweiser, R., Blouin, V., Toublanc, E. et al., A versatile and scalable two-step ion-exchange chromatography process for the purification of recombinant adeno-associated virus serotypes-2 and -5. Mol. Ther. 2002, 6, 678–686.
76. Okada, T., Nonaka-Sarukawa, M., Uchibori, R., Kinoshita, K. et al., Scalable purification of adeno-associated virus serotype 1 (AAV1) and AAV8 vectors, using dual ion-exchange adsorptive membranes. Hum. Gene Ther. 2009, 20, 1013–1021.
77. Urabe, M., Xin, K. Q., Obara, Y., Nakakura, T. et al., Removal of empty capsids from type 1 adeno-associated virus vector stocks by anion-exchange chromatography potentiates transgene expression. Mol. Ther. 2006, 13, 823–828.
78. Lock, M., Alvira, M. R., Wilson, J. M., Analysis of particle content of recombinant adeno-associated virus serotype 8 vectors by ion-exchange chromatography. Hum Gene Ther Methods 2012, 23, 56–64.
79. Qiu, J., Handa, a, Kirby, M., Brown, K. E., The interaction of heparin sulfate and adeno-associated virus 2. Virology 2000, 269, 137–147.
80. Halbert, C. L., Allen, J. M., Miller, A. D., Adeno-associated virus type 6 (AAV6) vectors mediate efficient transduction of airway epithelial cells in mouse lungs compared to that of AAV2 vectors. J. Virol. 2001, 75, 6615–6624.
81. Rabinowitz, J. E., Rolling, F., Li, C., Conrath, H. et al., Cross-packaging of a single adeno-associated virus (AAV) type 2 vector genome into multiple AAV serotypes enables transduction with broad specificity. J. Virol. 2002, 76, 791–801.
82. Liu, J., Moon, Y. A., Simple purification of adeno-associated virus-DJ for liver-specific gene expression. Yonse. Med. J. 2016, 57, 790–794.
83. Kaludov, N., Brown, K. E., Walters, R. W., Zabner, J., Chiorini, J. A., Adeno-associated virus serotype 4 (AAV4) and AAV5 both require sialic acid binding for hemagglutination and efficient transduction but differ in sialic acid linkage specificity. J. Virol. 2001, 75, 6884–6893.
84. Auricchio, A., A Single-step affinity column for purification of serotype-5 based adeno-associated viral vectors. Mol. Ther. 2001, 4, 372–374.
85. Wu, Z. J., Miller, E., Agbandje-McKenna, M., Samulski, R. J., alpha 2,3 and alpha 2,6 N-linked sialic acids facilitate efficient binding and transduction by adeno-associated virus types 1 and 6. J. Virol. 2006, 80, 9093–9103.
86. Smith, R. H., Levy, J. R., Kotin, R. M., A simplified baculovirus-AAV expression vector system coupled with one-step affinity purification yields high-titer rAAV stocks from insect cells. Mol. Ther. 2009, 17, 1888–1896.
87. Ayuso, E., Mingozzi, F., Montane, J., Leon, X. et al., High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency. Gene Ther. 2010, 17, 503–510.
88. Luchsinger, E., Strobbe, R., Dekegel, D., Wellemans, G., Use of B-IV zonal rotor centrifugation as a simple tool for the separation of adeno-associated X7 virus (AAVX7) from helper adenoviruses. Arch. Gesamte Virusforsch. 1971, 33, 251–258.
89. Peck, F. B., Purified influenza virus vaccine. A study of viral reactivity and antigenicity. JAMA 1968, 206, 2277–2282.
90. Brass, O., Contzen, J.-M., Francon, A., Tardy, M., Method for purifying a viral suspension. US-Patent US8016120 B2, 2011.
91. Merten, O.-W., Schweizer, M., Chahal, P., Kamen, A., Production and purification of viral vectors and safety considerations related to their use, in: Scherman, D. (Ed.), Advanced Textbook on Gene Transfer, Gene Therapy and Genetic Pharmacology: Principles, Delivery and Pharmacological and Biomedical Applications of Nucleotide-Based Therapies, World Scientific, Singapore 2013.
92. Martino, A. T., Suzuki, M., Markusic, D. M., Zolotukhin, I. et al., The genome of self-complementary adeno-associated viral vectors increases Toll-like receptor 9 – dependent innate immune responses in the liver. Gene Expr. 2012, 117, 6459–6468.
93. Faust, S. M., Bell, P., Cutler, B. J., Ashley, S. N. et al., CpG-depleted adeno-associated virus vectors evade immune detection. J. Clin. Invest. 2013, 123, 2994–3001.
94. Allay, J. A., Sleep, S., Long, S., Tillman, D. M. et al., Good manufacturing practice production of self-complementary serotype 8 adeno-associated viral vector for a hemophilia B clinical trial. Hum. Gene Ther. 2011, 22, 595–604.
95. Chadeuf, G., Ciron, C., Moullier, P., Salvetti, A., Evidence for encapsidation of prokaryotic sequences during recombinant adeno-associated virus production and their in vivo persistence after vector delivery. Mol. Ther. 2005, 12, 744–753.
96. Wright, J. F., Zelenaia, O., Vector characterization methods for quality control testing of recombinant adeno-associated viruses. Methods Mol. Biol. 2011, 737, 247–78.
97. Ye, G.-J., Scotti, M. M., Liu, J., Wang, L. et al., Clearance and characterization of residual HSV DNA in recombinant adeno-associated virus produced by an HSV complementation system. Gene Ther. 2011, 18, 135–144.
98. Meghrous, J., Aucoin, M. G., Jacob, D., Chahal, P. S. et al., Production of Recombinant Adeno-Associated Viral Vectors Using a Baculovirus / Insect Cell Suspension Culture System: From Shake Flasks to a 20-L Bioreactor. Biotechnol. Progr. 2005, 21, 154–160.
99. Nony, P., Chadeuf, G., Tessier, J., Moullier, P., Salvetti, A., Evidence for packaging of rep-cap sequences into adeno-associated virus (AAV) type 2 capsids in the absence of inverted terminal repeats: A model for generation of rep-positive AAV particles. J. Virol. 2003, 77, 776–781.
100. Hauck, B., Murphy, S. L., Smith, P. H., Qu, G. et al., Undetectable transcription of cap in a clinical AAV vector: Implications for preformed capsid in immune responses. Mol. Ther. 2009, 17, 144–152.
101. Lu, H., Qu, G., Yang, X., Xu, R., Xiao, W., Systemic elimination of de novo capsid protein synthesis from replication-competent AAV contamination in the liver. Hum. Gene Ther. 2011, 22, 625–632.
102. Lock, M., McGorray, S., Auricchio, A., Ayuso, E. et al., Characterization of a recombinant adeno-associated virus type 2 Reference Standard Material. Hum. Gene Ther. 2010, 21, 1273–1285.
103. Nathwani, A. C., Tuddenham, E. G. D., Rangarajan, S., Rosales, C. et al., Adenovirus-associated virus vector-mediated gene transfer in Hemophilia B. N. Engl. J. Med. 2011 365, 2357–2365.
104. Wright, J. F., AAV empty capsids: For better or for worse. Mol. Ther. 2014, 22, 1–2.
105. Grimm, D., Kern, A., Pawlita, M., Ferrari, F. et al., Titration of AAV-2 particles via a novel capsid ELISA: Packaging of genomes can limit production of recombinant AAV-2. Gene Ther. 1999, 6, 1322–1330.
106. Burnham, B., Nass, S., Kong, E., Mattingly, M. et al., Analytical ultracentrifugation as an approach to characterize recombinant adeno-associated viral vectors. Hum. Gene Ther. Methods 2015, 26, 228–242.
107. Pierson, E. E., Keifer, D. Z., Asokan, A., Jarrold, M. F., Resolving adeno-associated viral particle diversity with charge detection mass spectrometry. Anal. Chem. 2016, 88, 6718–6725.
108. Sommer, J. M., Smith, P. H., Parthasarathy, S., Isaacs, J. et al., Quantification of adeno-associated virus particles and empty capsids by optical density measurement. Mol. Ther. 2003 7, 122–128.
109. Kohlbrenner, E., Henckaerts, E., Rapti, K., Gordon, R. E. et al., Quantification of AAV particle titers by infrared fluorescence scanning of coomassie-stained sodium dodecyl sulfate-polyacrylamide gels. Hum. Gene Ther. Methods 2012, 23, 198–203.
110. Samulski, R. J., Chang, L. S., Shenk, T., Helper-free stocks of recombinant adeno-associated viruses: Normal integration does not require viral gene expression. J. Virol. 1989 63, 3822–3828.
111. Clark, K. R., Liu, X., McGrath, J. P., Johnson, P. R., Highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses. Hum. Gene Ther. 1999, 10, 1031–1039.
112. Fagone, P., Wright, J. F., Nathwani, A. C., Nienhuis, A. W. et al., Systemic errors in quantitative polymerase chain reaction titration of self-complementary adeno-associated viral vectors and improved alternative methods. Hum. Gene Ther. Methods 2012, 23, 1–7.
113. Pinheiro, L. B., Coleman, V. A., Hindson, C. M., Herrmann, J. et al., Evaluation of a droplet digital polymerase chain reaction format for DNA copy number quantification. Anal. Chem. 2012, 84, 1003–1011.
114. Lock, M., Alvira, M. R., Chen, S.-J., Wilson, J. M., Absolute determination of single-stranded and self-complementary adeno-associated viral vector genome titers by droplet digital PCR. Hum. Gene Ther. Methods 2014, 25, 115–125.
115. Baer, A., Kehn-Hall, K., Viral concentration determination through plaque assays: Using traditional and novel overlay systems. J. Vis. Exp. 2014, 4, e52065.
116. Lecomte, E., Tournaire, B., Cogné, B., Dupont, J.-B. et al., Advanced characterization of DNA molecules in rAAV vector preparations by single-stranded virus next-generation sequencing. Mol. Ther. Acids 2015, 4, e260.
117. Dong, B., Duan, X., Chow, H. Y., Chen, L. et al., Proteomics analysis of co-purifying cellular proteins associated with rAAV vectors. PLoS One 2014, 9, e86453.
118. Sommer, J. M., Smith, P. H., Parthasarathy, S., Isaacs, J. et al., Quantification of adeno-associated virus particles and empty capsids by optical density measurement. Mol. Ther. 2003, 7, 122–128.
119. Grimm, D., Kay, M. A., Kleinschmidt, J. A., Helper virus-free, optically controllable, and two-plasmid-based production of adeno-associated virus vectors of serotypes 1 to 6. Mol. Ther. 2003, 7, 839–850.
120. Wright, J. F., Manufacturing and characterizing AAV-based vectors for use in clinical studies. Gene Ther. 2008, 15, 840–848.
121. Mitchell, D. A. J., Lerch, T. F., Hare, J. T., Chapman, M. S., A pseudo-plaque method for infectious particle assay and clonal isolation of adeno-associated virus. J. Virol. Methods 2010, 170, 9–15.
122. Zen, Z., Espinoza, Y., Bleu, T., Sommer, J. M., Wright, J. F., Infectious titer assay for adeno-associated virus vectors with sensitivity sufficient to detect single infectious events. Hum. Gene Ther. 2004, 15, 709–715.
123. Rohr, U.-P., Wulf, M.-A., Stahn, S., Steidl, U. et al., Fast and reliable titration of recombinant adeno-associated virus type-2 using quantitative real-time PCR. J. Virol. Methods 2002, 106, 81–88.
124. Piedra, J., Ontiveros, M., Miravet, S., Penalva, C. et al., Development of a rapid, robust, and universal PicoGreen-based method to titer adeno-associated vectors. Hum. Gene Ther. Methods 2015, 26, 35–42.
125. Halbert, C. L., Standaert, T. A., Aitken, M. L., Alexander, I. E. et al., Transduction by adeno-associated virus vectors in the rabbit airway: Efficiency, persistence, and readministration. J. Virol. 1997, 71, 5932–5941.
126. Horowitz, E. D., Rahman, K. S., Bower, B. D., Dismuke, D. J. et al., Biophysical and ultrastructural characterization of adeno-associated virus capsid uncoating and genome release. J. Virol. 2013, 87, 2994–3002.
127. Kronenberg, S., Kleinschmidt, J. A., Böttcher, B., Electron cryo-microscopy and image reconstruction of adeno-associated virus type 2 empty capsids. EMBO Rep, 2001, 2, 997–1002.
128. Kleibeuker, W., Zhou, X., Centlivre, M., Legrand, N. et al., A sensitive cell-based assay to measure the doxycycline concentration in biological samples. Hum. Gene Ther. 2009, 20, 524–530.
129. Krishna, Ac., Sathiyaraj, M., Saravanan, R., Chelladurai, R., Vignesh, R., A novel and rapid method to determine doxycycline in human plasma by liquid chromatography tandem mass spectrometry. Indian J. Pharm. Sci. 2012, 74, 541–548.
130. Hermens, W. T., ter Brake, O., Dijkhuizen, P. A., Sonnemans, M. A. et al., Purification of recombinant adeno-associated virus by iodixanol gradient ultracentrifugation allows rapid and reproducible preparation of vector stocks for gene transfer in the nervous system. Hum. Gene Ther. 1999, 10, 1885–1891.
131. Wang, Z., Storb, R., Halbert, C. L., Banks, G. B. et al., Successful regional delivery and long-term expression of a dystrophin gene in canine muscular dystrophy: A preclinical model for human therapies. Mol. Ther. 2012, 20, 1501–1507.
132. Bharadwaj, A. S., Kelly, M., Kim, D., Chao, H., Induction of immune tolerance to FIX by intramuscular AAV gene transfer is independent of the activation status of dendritic cells. Blood 2010, 115, 500–509.
133. Wendtner, C.-M. M., Kofler, D. M., Theiss, H. D., Kurzeder, C. et al., Efficient gene transfer of CD40 ligand into primary B-CLL cells using recombinant adeno-associated virus (rAAV) vectors. Blood 2002, 100, 1655–1661.
134. Volokhov, D. V., Graham, L. J., Brorson, K. A., Chizhikov, V. E., Mycoplasma testing of cell substrates and biologics: Review of alternative non-microbiological techniques. Mol. Cell. Probes 2011, 25, 69–77.
135. Flotte, T. R., Trapnell, B. C., Humphries, M., Carey, B. et al., Phase 2 clinical trial of a recombinant adeno-associated viral vector expressing α1-antitrypsin: Interim results. Hum. Gene Ther. 2011, 22, 1239–1247.
136. Mingozzi, F., High, K. A., Immune responses to AAV vectors: Overcoming barriers to successful gene therapy. Blood 2013, 122, 23–36.
137. Ding, J. L., Ho, B., Endotoxin detection – from limulus amebocyte lysate to recombinant factor C. Subcell. Biochem. 2010, 53, 187–208.
138. Salganik, M., Venkatakrishnan, B., Bennett, A., Lins, B. et al., Evidence for pH-dependent protease activity in the adeno-associated virus capsid. J. Virol. 2012, 86, 11877–11885.