Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material

Eduard Ayuso; Véronique Blouin; Martin Lock; Susan Mcgorray; Xavier Leon; Mauricio R. Alvira; Alberto Auricchio; Stephanie Bucher; Abdelwahed Chtarto; K. Reed Clark; Christophe Darmon; Monica Doria; Will Fountain; Guangping Gao; Kai Gao; Mauro Giacca; Juergen Kleinschmidt; Barbara Leuchs; Catherine Melas; Hiroaki Mizukami; Marcus Müller; Yvet Noordman; Olivier Bockstael; Keiya Ozawa; Catherine Pythoud; Marina Sumaroka; Richard Surosky; Liliane Tenenbaum; Inge Van Der Linden; Brigitte Weins; J. Fraser Wright; Xinhua Zhang; Lorena Zentilin; Fatima Bosch; Richard O. Snyder; Philippe Moullier

doi:10.1089/hum.2014.057

Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material

Eduard Ayuso, Véronique Blouin, Martin Lock, Susan Mcgorray, Xavier Leon, Mauricio R. Alvira, Alberto Auricchio, Stephanie Bucher, Abdelwahed Chtarto, K. Reed Clark, Christophe Darmon, Monica Doria, Will Fountain, Guangping Gao, Kai Gao, Mauro Giacca, Juergen Kleinschmidt, Barbara Leuchs, Catherine Melas, Hiroaki MizukamiMarcus Müller, Yvet Noordman, Olivier Bockstael, Keiya Ozawa, Catherine Pythoud, Marina Sumaroka, Richard Surosky, Liliane Tenenbaum, Inge Van Der Linden, Brigitte Weins, J. Fraser Wright, Xinhua Zhang, Lorena Zentilin, Fatima Bosch, Richard O. Snyder, Philippe Moullier^*

^*Corresponding author for this work

Cardiovascular Medicine & Sciences

Research output: Contribution to journal › Article › peer-review

80 Citations (Scopus)

Abstract

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, 5.50×10¹¹ pt/ml; CI, 4.26×10¹¹ to 6.75×10¹¹ pt/ml), vector genomes (mean, 5.75×10¹¹ vg/ml; CI, 3.05×10¹¹ to 1.09×10¹² vg/ml), and infectious units (mean, 1.26×10⁹ IU/ml; CI, 6.46×10⁸ to 2.51×10⁹ 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.

Original language	English
Pages (from-to)	977-987
Number of pages	11
Journal	Human Gene Therapy
Volume	25
Issue number	11
DOIs	https://doi.org/10.1089/hum.2014.057
Publication status	Published - 1 Jan 2014

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1089/hum.2014.057

Cite this

Ayuso, E., Blouin, V., Lock, M., Mcgorray, S., Leon, X., Alvira, M. R., Auricchio, A., Bucher, S., Chtarto, A., Clark, K. R., Darmon, C., Doria, M., Fountain, W., Gao, G., Gao, K., Giacca, M., Kleinschmidt, J., Leuchs, B., Melas, C., ... Moullier, P. (2014). Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material. Human Gene Therapy, 25(11), 977-987. https://doi.org/10.1089/hum.2014.057

@article{5c8a2718a5454183b009b87304bf1532,

title = "Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material",

abstract = "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, 5.50×1011 pt/ml; CI, 4.26×1011 to 6.75×1011 pt/ml), vector genomes (mean, 5.75×1011 vg/ml; CI, 3.05×1011 to 1.09×1012 vg/ml), and infectious units (mean, 1.26×109 IU/ml; CI, 6.46×108 to 2.51×109 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.",

author = "Eduard Ayuso and V{\'e}ronique Blouin and Martin Lock and Susan Mcgorray and Xavier Leon and Alvira, {Mauricio R.} and Alberto Auricchio and Stephanie Bucher and Abdelwahed Chtarto and Clark, {K. Reed} and Christophe Darmon and Monica Doria and Will Fountain and Guangping Gao and Kai Gao and Mauro Giacca and Juergen Kleinschmidt and Barbara Leuchs and Catherine Melas and Hiroaki Mizukami and Marcus M{\"u}ller and Yvet Noordman and Olivier Bockstael and Keiya Ozawa and Catherine Pythoud and Marina Sumaroka and Richard Surosky and Liliane Tenenbaum and {Van Der Linden}, Inge and Brigitte Weins and Wright, {J. Fraser} and Xinhua Zhang and Lorena Zentilin and Fatima Bosch and Snyder, {Richard O.} and Philippe Moullier",

year = "2014",

month = jan,

day = "1",

doi = "10.1089/hum.2014.057",

language = "English",

volume = "25",

pages = "977--987",

journal = "Human Gene Therapy",

issn = "1043-0342",

publisher = "Mary Ann Liebert Inc.",

number = "11",

}

Ayuso, E, Blouin, V, Lock, M, Mcgorray, S, Leon, X, Alvira, MR, Auricchio, A, Bucher, S, Chtarto, A, Clark, KR, Darmon, C, Doria, M, Fountain, W, Gao, G, Gao, K, Giacca, M, Kleinschmidt, J, Leuchs, B, Melas, C, Mizukami, H, Müller, M, Noordman, Y, Bockstael, O, Ozawa, K, Pythoud, C, Sumaroka, M, Surosky, R, Tenenbaum, L, Van Der Linden, I, Weins, B, Wright, JF, Zhang, X, Zentilin, L, Bosch, F, Snyder, RO & Moullier, P 2014, 'Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material', Human Gene Therapy, vol. 25, no. 11, pp. 977-987. https://doi.org/10.1089/hum.2014.057

TY - JOUR

T1 - Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material

AU - Ayuso, Eduard

AU - Blouin, Véronique

AU - Lock, Martin

AU - Mcgorray, Susan

AU - Leon, Xavier

AU - Alvira, Mauricio R.

AU - Auricchio, Alberto

AU - Bucher, Stephanie

AU - Chtarto, Abdelwahed

AU - Clark, K. Reed

AU - Darmon, Christophe

AU - Doria, Monica

AU - Fountain, Will

AU - Gao, Guangping

AU - Gao, Kai

AU - Giacca, Mauro

AU - Kleinschmidt, Juergen

AU - Leuchs, Barbara

AU - Melas, Catherine

AU - Mizukami, Hiroaki

AU - Müller, Marcus

AU - Noordman, Yvet

AU - Bockstael, Olivier

AU - Ozawa, Keiya

AU - Pythoud, Catherine

AU - Sumaroka, Marina

AU - Surosky, Richard

AU - Tenenbaum, Liliane

AU - Van Der Linden, Inge

AU - Weins, Brigitte

AU - Wright, J. Fraser

AU - Zhang, Xinhua

AU - Zentilin, Lorena

AU - Bosch, Fatima

AU - Snyder, Richard O.

AU - Moullier, Philippe

PY - 2014/1/1

Y1 - 2014/1/1

N2 - 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, 5.50×1011 pt/ml; CI, 4.26×1011 to 6.75×1011 pt/ml), vector genomes (mean, 5.75×1011 vg/ml; CI, 3.05×1011 to 1.09×1012 vg/ml), and infectious units (mean, 1.26×109 IU/ml; CI, 6.46×108 to 2.51×109 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.

AB - 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, 5.50×1011 pt/ml; CI, 4.26×1011 to 6.75×1011 pt/ml), vector genomes (mean, 5.75×1011 vg/ml; CI, 3.05×1011 to 1.09×1012 vg/ml), and infectious units (mean, 1.26×109 IU/ml; CI, 6.46×108 to 2.51×109 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.

UR - http://www.scopus.com/inward/record.url?scp=84910600504&partnerID=8YFLogxK

U2 - 10.1089/hum.2014.057

DO - 10.1089/hum.2014.057

M3 - Article

C2 - 25275822

AN - SCOPUS:84910600504

SN - 1043-0342

VL - 25

SP - 977

EP - 987

JO - Human Gene Therapy

JF - Human Gene Therapy

IS - 11

ER -

Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material

Abstract

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this