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Production and Formulation of Adenovirus Vectors

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Gene Therapy and Gene Delivery Systems

Part of the book series: Advances in Biochemical Engineering/Biotechnology ((ABE,volume 99))

Abstract

Adenovirus vectors have attracted considerable interest over the past decade, with ongoing clinical development programs for applications ranging from replacement therapy for protein deficiencies to cancer therapeutics to prophylactic vaccines. Consequently, considerable product, process, analytical, and formulation development has been undertaken to support these programs. For example, “gutless” vectors have been developed in order to improve gene transfer capacity and durability of expression; new cell lines have been developed to minimize recombination events; production conditions have been optimized to improve volumetric productivities; analytical techniques and scaleable purification processes have advanced towards the goal of purified adenovirus becoming a “well-characterized biological”; and liquid formulations have been developed which maintain virus infectivity at 2–8°C for over 18 months. These and other advances in the production of adenovirus vectors are discussed in detail in this review. In addition, the needs for the next decade are highlighted.

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Abbreviations

AdV:

adenovirus

Ad2:

adenovirus type 2

Ad5:

adenovirus type 5

AEX:

anion exchange

ARM:

adenovirus reference material

CPE:

cytopathic effect

DELFIA:

dissociation-enhanced flouroimmunoassay

ELISA:

enzyme-linked immunosorbent assay

GMP:

good manufacturing practice

hdAdV:

helper-dependent adenovirus

HDEP:

helper-dependent E1-positive particle

hpi:

hours post-infection

MOI:

multiplicity of infection

P/I:

particle-to-infectivity ratio

Pfu:

plaque-forming unit

RCA:

replication-competent adenovirus

rpHPLC:

reversed-phase high performance liquid chromatography

RSD:

relative standard deviation

TCID50 :

tissue culture infectious dose at 50%

QPCR:

quantitative polymerase chain reaction

vp:

viral particles

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Acknowledgments

We thank coworkers who contributed to the generation of unpublished data presented in this work (Fig. 2: Marie-Pierre Gentile and Gina Cremona; Fig. 4: Matthew Kalscheur, Marie-Pierre Gentile, and Benjamin Hughes; Figs. 8 and 9: Denise Nawrocki, Lynne Isopi, and Donna Williams) and Drs. Angelica Meneses-Acosta and Olivier Henry for providing data and assistance with Figs. 5 and 6. We also thank Dr. John Lewis and Dr. Katey Einterz Owen for reviewing portions of this manuscript and providing helpful suggestions.

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Authors and Affiliations

  1. Fermentation and Cell Culture, Merck Research Laboratories, Sumneytown Pike, Pennsylvannia 19486-0004, West Point, USA

    Nedim E. Altaras & John G. Aunins

  2. Vaccine Pharmaceutical Research, Merck Research Laboratories, Sumneytown Pike, Pennsylvannia 19486-0004, West Point, USA

    Robert K. Evans

  3. Biologics Development and Engineering, Merck Research Laboratories, Sumneytown Pike, Pennsylvannia 19486-0004, West Point, USA

    John O. Konz

  4. Live Viral Vectors, Merck Research Laboratories, Sumneytown Pike, Pennsylvannia 19486-0004, West Point, USA

    Jayanthi J. Wolf

  5. Biotech Research Institute, National Research Council, 6100 Royalmount Avenue, H4P 2R2, Montreal, Quebec, Canada

    Amine Kamen

  6. VaxInnate, 3 Cedar Brook Dr., New Jersey 08512, Cranbury, USA

    Robert K. Evans

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  1. Nedim E. Altaras
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  2. John G. Aunins
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Correspondence to John O. Konz .

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Altaras, N.E., Aunins, J.G., Evans, R.K., Kamen, A., Konz, J.O., Wolf, J.J. Production and Formulation of Adenovirus Vectors. In: Gene Therapy and Gene Delivery Systems. Advances in Biochemical Engineering/Biotechnology, vol 99. Springer, Berlin, Heidelberg. https://doi.org/10.1007/10_008

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