Manufacturing and Characterization of a Recombinant Adeno-Associated Virus Type 8 Reference Standard Material

被引：71

作者：

Ayuso, Eduard ^{[1
,2
]}

Blouin, Veronique ^{[3
,4
]}

Lock, Martin ^{[5
]}

McGorray, Susan ^{[6
,7
]}

Leon, Xavier ^{[1
,2
]}

Alvira, Mauricio R. ^{[5
]}

Auricchio, Alberto ^{[8
,9
]}

Bucher, Stephanie ^{[10
]}

Chtarto, Abdelwahed ^{[11
,12
]}

Clark, K. Reed ^{[13
,14
]}

Darmon, Christophe ^{[3
,4
,15
]}

Doria, Monica ^{[9
]}

Fountain, Will ^{[13
,14
]}

Gao, Guangping ^{[16
]}

Gao, Kai ^{[16
,17
]}

Giacca, Mauro ^{[18
]}

Kleinschmidt, Juergen ^{[19
]}

Leuchs, Barbara ^{[19
]}

Melas, Catherine ^{[11
,12
]}

Mizukami, Hiroaki ^{[20
]}

Mueller, Marcus ^{[19
]}

Noordman, Yvet ^{[21
]}

Bockstael, Olivier ^{[11
,12
]}

Ozawa, Keiya ^{[20
]}

Pythoud, Catherine ^{[22
,23
]}

Sumaroka, Marina ^{[24
]}

Surosky, Richard ^{[25
]}

Tenenbaum, Liliane ^{[22
,23
]}

van der Linden, Inge ^{[21
]}

Weins, Brigitte ^{[26
]}

Wright, J. Fraser ^{[24
,27
]}

Zhang, Xinhua ^{[24
]}

Zentilin, Lorena ^{[18
]}

Bosch, Fatima ^{[1
,2
]}

Snyder, Richard O. ^{[28
,29
,30
]}

Moullier, Philippe ^{[3
,4
,30
]}

机构：

[1] Univ Autonoma Barcelona, Sch Vet Med, Ctr Anim Biotechnol & Gene Therapy, Bellaterra 08193, Spain

[2] Univ Autonoma Barcelona, Sch Vet Med, Dept Biochem & Mol Biol, Bellaterra 08193, Spain

[3] CHU Nantes, INSERM, UMR1089, F-44000 Nantes, France

[4] Atlantic Gene Therapies, F-44000 Nantes, France

[5] Univ Penn, Dept Pathol & Lab Med, Gene Therapy Program, Philadelphia, PA 19104 USA

[6] Univ Florida, Dept Biostat, Coll Med & Publ Hlth, Gainesville, FL 32610 USA

[7] Univ Florida, Dept Biostat, Coll Hlth Profess, Gainesville, FL 32610 USA

[8] Univ Naples Federico II, Dept Translat Med, I-80131 Naples, Italy

[9] Telethon Inst Genet & Med, TIGEM, I-80131 Naples, Italy

[10] GENETHON, F-91000 Evry, France

[11] Free Univ Brussels, Lab Expt Neurosurg, B-1070 Brussels, Belgium

[12] Free Univ Brussels, Multidisciplinary Res Inst, B-1070 Brussels, Belgium

[13] Nationwide Childrens Hosp, Ctr Gene Therapy, Res Inst, Columbus, OH 43205 USA

[14] Ohio State Univ, Dept Pediat, Columbus, OH 43210 USA

[15] EFS Atlantic BioGMP, F-44800 Nantes, France

[16] Univ Massachusetts, Sch Med, Gene Therapy Ctr, Worcester, MA 01605 USA

[17] Natl Inst Food & Drug Control, Div Biopharmaceut, Beijing 100050, Peoples R China

[18] ICGEB, Mol Med Lab, I-34149 Trieste, Italy

[19] German Canc Res Ctr, Res Program Infect & Canc, D-69120 Heidelberg, Germany

[20] Jichi Med Univ, Ctr Mol Med, Div Genet Therapeut, Shimotsuke, Tochigi 3290498, Japan

[21] uniQure, NL-1100 Amsterdam, Netherlands

[22] Univ Lausanne Hosp, Dept Clin Neurosci, Lab Cellular & Mol Neurotherapies, CH-1015 Lausanne, Switzerland

[23] Univ Lausanne Hosp, Dept Clin Neurosci, Neurosurg Serv, CH-1015 Lausanne, Switzerland

[24] Childrens Hosp Philadelphia, Ctr Cellular & Mol Therapeut, Philadelphia, PA 19104 USA

[25] Sangamo Biosci Inc, Richmond, CA 94804 USA

[26] Progen Biotech GmbH, D-69123 Heidelberg, Germany

[27] Univ Penn, Sch Med, Dept Pathol & Lab Med, Philadelphia, PA 19104 USA

[28] Univ Florida, Ctr Excellence Regenerat Hlth Biotechnol, Gainesville, FL 32611 USA

[29] Univ Florida, Dept Pediat, Div Cellular & Mol Therapy, Gainesville, FL 32610 USA

[30] Univ Florida, Coll Med, Dept Mol Genet & Microbiol, Gainesville, FL 32610 USA

来源：

HUMAN GENE THERAPY | 2014年 / 25卷 / 11期

关键词：

LEBERS CONGENITAL AMAUROSIS; VECTOR REFERENCE-STANDARDS; GENE-TRANSFER; VIRAL VECTORS; TRANSDUCTION; HEMOPHILIA; EFFICIENCY; PARTICLES; THERAPY; CAPSIDS;

D O I：

10.1089/hum.2014.057

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Gene therapy approaches using recombinant adeno-associated virus serotype 2 (rAAV2) and serotype 8 (rAAV8) have achieved significant clinical benefits. The generation of rAAV Reference Standard Materials (RSM) is key to providing points of reference for particle titer, vector genome titer, and infectious titer for gene transfer vectors. Following the example of the rAAV2RSM, here we have generated and characterized a novel RSM based on rAAV serotype 8. The rAAV8RSM was produced using transient transfection, and the purification was based on density gradient ultracentrifugation. The rAAV8RSM was distributed for characterization along with standard assay protocols to 16 laboratories worldwide. Mean titers and 95% confidence intervals were determined for capsid particles (mean, 5.50 x 10(11) pt/ml; CI, 4.26 x 10(11) to 6.75 x 10(11) pt/ml), vector genomes (mean, 5.75 x 10(11) vg/ml; CI, 3.05 x 10(11) to 1.09 x 10(12) vg/ml), and infectious units (mean, 1.26 x 10(9) IU/ml; CI, 6.46 x 10(8) to 2.51 x 10(9) IU/ml). Notably, there was a significant degree of variation between institutions for each assay despite the relatively tight correlation of assay results within an institution. This outcome emphasizes the need to use RSMs to calibrate the titers of rAAV vectors in preclinical and clinical studies at a time when the field is maturing rapidly. The rAAV8RSM has been deposited at the American Type Culture Collection (VR-1816) and is available to the scientific community.

引用

页码：977 / 987

页数：11

共 33 条

[1] High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency [J].

Ayuso, E. ;

Mingozzi, F. ;

Montane, J. ;

Leon, X. ;

Anguela, X. M. ;

Haurigot, V. ;

Edmonson, S. A. ;

Africa, L. ;

Zhou, S. ;

High, K. A. ;

Bosch, F. ;

Wright, J. F. .

GENE THERAPY, 2010, 17 (04) :503-510

[2]

Ayuso E., 2012, CLINIBOOK CLIN GENE, P83

[3] Effect of gene therapy on visual function in Leber's congenital amaurosis [J].

Bainbridge, James W. B. ;

Smith, Alexander J. ;

Barker, Susie S. ;

Robbie, Scott ;

Henderson, Robert ;

Balaggan, Kamaljit ;

Viswanathan, Ananth ;

Holder, Graham E. ;

Stockman, Andrew ;

Tyler, Nick ;

Petersen-Jones, Simon ;

Bhattacharya, Shomi S. ;

Thrasher, Adrian J. ;

Fitzke, Fred W. ;

Carter, Barrie J. ;

Rubin, Gary S. ;

Moore, Anthony T. ;

Ali, Robin R. .

NEW ENGLAND JOURNAL OF MEDICINE, 2008, 358 (21) :2231-2239

[4]

Bryant LR, 2013, PERS CONTIN PHILOS, P55

[5] Recombinant AAV viral vectors pseudotyped with viral capsids from serotypes 1, 2, and 5 display differential efficiency and cell tropism after delivery to different regions of the central nervous system [J].

Burger, C ;

Gorbatyuk, OS ;

Velardo, MJ ;

Peden, CS ;

Williams, P ;

Zolotukhin, S ;

Reier, PJ ;

Mandel, RJ ;

Muzyczka, N .

MOLECULAR THERAPY, 2004, 10 (02) :302-317

[6] Clades of Adeno-associated viruses are widely disseminated in human tissues [J].

Gao, GP ;

Vandenberghe, LH ;

Alvira, MR ;

Lu, Y ;

Calcedo, R ;

Zhou, XY ;

Wilson, JA .

JOURNAL OF VIROLOGY, 2004, 78 (12) :6381-6388

[7] Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy [J].

Gao, GP ;

Alvira, MR ;

Wang, LL ;

Calcedo, R ;

Johnston, J ;

Wilson, JM .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (18) :11854-11859

[8] Efficacy and long-term safety of alipogene tiparvovec (AAV1-LPLS447X) gene therapy for lipoprotein lipase deficiency: an open-label trial [J].

Gaudet, D. ;

Methot, J. ;

Dery, S. ;

Brisson, D. ;

Essiembre, C. ;

Tremblay, G. ;

Tremblay, K. ;

de Wal, J. ;

Twisk, J. ;

van den Bulk, N. ;

Sier-Ferreira, V. ;

van Deventer, S. .

GENE THERAPY, 2013, 20 (04) :361-369

[9] Titration of AAV-2 particles via a novel capsid ELISA: packaging of genomes can limit production of recombinant AAV-2 [J].

Grimm, D ;

Kern, A ;

Pawlita, M ;

Ferrari, FK ;

Samulski, RJ ;

Kleinschmidt, JA .

GENE THERAPY, 1999, 6 (07) :1322-1330

[10]

Hutchins B., 2002, Bioprocess. J, V1, P25, DOI [10.12665/J11.Hutchins, DOI 10.12665/J11, DOI 10.12665/J11.HUTCHINS]

← 1 2 3 4 →