Skip to main content

Epitope Binning of Human Monoclonal Antibodies in Classical Sandwich and In-Tandem Orientation Using the Octet System Based on Biolayer Interferometry

  • Protocol
  • First Online:
Epitope Mapping Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1785))

Abstract

Epitope binning is a technique used to cluster different monoclonal antibodies by the specific region on the antigen that is recognized by the antibody. This is used to increase the likelihood of choosing lead antibodies with the desired biological activity by selecting antibodies from distinct bins, to confirm that biosimilars or biobetters bind to similar epitopes and to select reagents for diagnostic sandwich and/or ELISA-type assays. This protocol describes epitope binning of seven human monoclonal antibodies to a monovalent antigen in two orthogonal orientations; a classical sandwich and an in-tandem orientation using an eight-channel Octet system.

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

Access this chapter

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 199.99
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. Dong J, Demarest S, Sereno A et al (2010) Combination of two insulin-like growth factor-I receptor inhibitory antibodies targeting distinct epitopes leads to an enhanced antitumor response. Mol Cancer Ther 9:2593–2604

    Article  CAS  PubMed  Google Scholar 

  2. Koefoed K, Steinaa L, Soderberg L et al (2011) Rational indentiication of an optimal anitbody mixture for targeting the epidermal growth factor receptor. MAbs 3:584–595

    Article  PubMed Central  PubMed  Google Scholar 

  3. Niederfellner G, Lammens A, Mudigl O et al (2011) Epitope characterization and chrsytal structure of GA101 provide insights into the molecular basis for type I/II distinction of CD20 antibodies. Blood 118:358–367

    Article  CAS  PubMed  Google Scholar 

  4. Byrd J, Kipps T, Flinn I et al (2012) Phase I study of the anti-CD40 humanized monoclonal anitbody lucatumumab (HCD122) in relapsed chronic lymphocytic leukemia. Leuk Lymphoma 53:2136–2142

    Article  CAS  PubMed  Google Scholar 

  5. Hellstrom K, Yelton D, Fell H et al (1990) Epitope mapping and use of anti-idiotypic antibodies to the L6 monoclonal anticarcinoma antibody. Cancer Res 50:2449–2454

    CAS  PubMed  Google Scholar 

  6. Abdiche Y, Lindquist K, Stone D et al (2012) Label-free epitope binning assays of monoclonal antibodies enable identification of antigen heterogeneity. J Immunol Methods 382:101–116

    Article  CAS  PubMed  Google Scholar 

  7. Abdiche Y, Malashock D, Pinkerton A et al (2009) Exploring blocking assays using octet, ProteOn, and Biacore biosensors. Anal Biochem 386:172–180

    Article  CAS  PubMed  Google Scholar 

  8. Takkar R, Yadav A, Kumaraswamy S et al (2013) Cross-competition or epitope binning assays on the octet HTX system. Application Note of Pall Life Sciences, Menlo Park

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lisette Bronswijk-Deddens .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Bronswijk-Deddens, L. (2018). Epitope Binning of Human Monoclonal Antibodies in Classical Sandwich and In-Tandem Orientation Using the Octet System Based on Biolayer Interferometry. In: Rockberg, J., Nilvebrant, J. (eds) Epitope Mapping Protocols. Methods in Molecular Biology, vol 1785. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7841-0_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7841-0_13

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7839-7

  • Online ISBN: 978-1-4939-7841-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics