Skip to main content
SpringerLink
Log in

Production, Purification, and Titration of First-Generation Adenovirus Vectors

  • Protocol
  • First Online:
Functional Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1654))

Abstract

Vectors based on human adenovirus are highly efficient tools for transient genetic modifications of cells or tissues in vitro and in vivo. They can be utilized for gene addition strategies, knockdown strategies and as transfer vectors for designer nucleases and CRISPR/Cas. They are characterized by high genomic stability and can be produced to high titers. This chapter describes the method how to produce, purify and titrate adenovirus vectors based on human adenovirus type 5.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 179.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Volpers C, Kochanek S (2004) Adenoviral vectors for gene transfer and therapy. J Gene Med 6(Suppl 1):S164–S171. doi:10.1002/jgm.496

    Article  CAS  PubMed  Google Scholar 

  2. Graham FL, Smiley J, Russell WC, Nairn R (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36:59–74. doi:10.1099/0022-1317-36-1-59

    Article  CAS  PubMed  Google Scholar 

  3. Bergelson JM, Cunningham JA, Droguett G et al (1997) Isolation of a common receptor for Coxsackie B viruses and adenoviruses 2 and 5. Science 275:1320–1323

    Article  CAS  PubMed  Google Scholar 

  4. Wickham TJ, Mathias P, Cheresh DA, Nemerow GR (1993) Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell 73:309–319

    Article  CAS  PubMed  Google Scholar 

  5. Trotman LC, Mosberger N, Fornerod M et al (2001) Import of adenovirus DNA involves the nuclear pore complex receptor CAN/Nup214 and histone H1. Nat Cell Biol 3:1092–1100. doi:10.1038/ncb1201-1092

    Article  CAS  PubMed  Google Scholar 

  6. Maizel JV, White DO, Scharff MD (1968) The polypeptides of adenovirus: I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12. Virology 36:115–125

    Article  CAS  PubMed  Google Scholar 

  7. Mittereder N, March KL, Trapnell BC (1996) Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J Virol 70:7498–7509

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Kreppel F, Biermann V, Kochanek S, Schiedner G (2002) A DNA-based method to assay total and infectious particle contents and helper virus contamination in high-capacity adenoviral vector preparations. Hum Gene Ther 13:1151–1156. doi:10.1089/104303402320138934

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Gene Therapy, Ulm University, Helmholtzstraße 8/1, 89081, Ulm, Germany

    Ramona F. Kratzer

  2. Chair of Biochemistry and Molecular Medicine, Faculty of Health, School of Medicine, Center for Biomedical Research and Education ZBAF, Witten/Herdecke University, Stockumer Straße 10, 58453, Witten, Germany

    Florian Kreppel

Authors
  1. Ramona F. Kratzer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Florian Kreppel
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Florian Kreppel .

Editor information

Editors and Affiliations

  1. Chair of Biochemistry and Molecular Medicine, Division of Functional Genomics, Faculty of Health, School of Medicine, Center for Biochemical Research and Education ZBAF, Witten/Herdecke University, Witten, Germany

    Michael Kaufmann

  2. Chair of Biochemistry and Molecular Medicine, Division of Functional Genomics, Faculty of Health, School of Medicine, Center for Biochemical Research and Education ZBAF, Witten/Herdecke University, Witten, Germany

    Claudia Klinger

  3. Chair of Biochemistry and Molecular Medicine, Division of Functional Genomics, Faculty of Health, School of Medicine, Center for Biochemical Research and Education ZBAF, Witten/Herdecke University, Witten, Germany

    Andreas Savelsbergh

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Kratzer, R.F., Kreppel, F. (2017). Production, Purification, and Titration of First-Generation Adenovirus Vectors. In: Kaufmann, M., Klinger, C., Savelsbergh, A. (eds) Functional Genomics. Methods in Molecular Biology, vol 1654. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7231-9_28

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7231-9_28

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7230-2

  • Online ISBN: 978-1-4939-7231-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation