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Functional Characterization of Antibodies Neutralizing Soluble Factors In Vitro and In Vivo

  • Geertruida M. VeldmanEmail author
  • Zehra Kaymakcalan
  • Renee Miller
  • Leena Kalghatgi
  • Jochen G. Salfeld
Protocol
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Part of the Springer Protocols Handbooks book series (SPH)

Functional characterization of antibodies that inhibit soluble cytokines or chemokines requires robust, sensitive in vitro and in vivo bioassays. Testing an antibody in vitro requires consideration of antigen source, integrity, and concentration, as well as the magnitude of the biologic response and assay interference by components in the antibody test sample. This chapter describes several exemplary in vitro bioassays, including an assay to determine species cross-reactivity of an anti–tumor necrosis factor antibody and a whole blood assay for interleukin–18 (IL–18) to determine neutralization of native antigen. Testing an antibody in vivo requires consideration of species cross-reactivity, selection of the dose range and the route of administration, analyses of the pharmacokinetics and tissue distribution of the antibody, and evaluation of host antibody responses. An example of an in vivo bioassay, in which human peripheral blood mononuclear cells are injected into severe combined immunodeficient mice and activated in vivo to produce human IL–18, is provided.

Keywords

Antigen Concentration L929 Culture Medium Human Tumor Necrosis Factor Native Antigen Antigen Source 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

Editorial support was provided by Robin L. Stromberg, PhD, of Arbor Communications, Inc. (Ann Arbor, MI, USA) and funded by Abbott Laboratories.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2010

Authors and Affiliations

  • Geertruida M. Veldman
    • 1
    Email author
  • Zehra Kaymakcalan
    • 1
  • Renee Miller
    • 1
  • Leena Kalghatgi
    • 1
  • Jochen G. Salfeld
    • 1
  1. 1.Department of BiologicsAbbott Bioresearch CenterWorcesterUSA

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