Cancer Immunology, Immunotherapy

, Volume 57, Issue 12, pp 1879–1890 | Cite as

Kinetics of anti-carcinoembryonic antigen antibody internalization: effects of affinity, bivalency, and stability

  • Michael M. Schmidt
  • Greg M. Thurber
  • K. Dane Wittrup
Original Article

Abstract

Theoretical analyses suggest that the cellular internalization and catabolism of bound antibodies contribute significantly to poor penetration into tumors. Here we quantitatively assess the internalization of antibodies and antibody fragments against the commonly targeted antigen carcinoembryonic antigen (CEA). Although CEA is often referred to as a non-internalizing or shed antigen, anti-CEA antibodies and antibody fragments are shown to be slowly endocytosed by LS174T cells with a half-time of 10–16 h, a time scale consistent with the metabolic turnover rate of CEA in the absence of antibody. Anti-CEA single chain variable fragments (scFvs) with significant differences in affinity, stability against protease digestion, and valency exhibit similar uptake rates of bound antibody. In contrast, one anti-CEA IgG exhibits unique binding and trafficking properties with twice as many molecules bound per cell at saturation and significantly faster cellular internalization after binding. The internalization rates measured herein can be used in simple computational models to predict the microdistribution of these antibodies in tumor spheroids.

Keywords

Tumor targeting CEA Endocytosis Antibody fragments Affinity 

Supplementary material

262_2008_518_MOESM1_ESM.pdf (1.2 mb)
MOESM1 (PDF 96 kb)

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Michael M. Schmidt
    • 1
  • Greg M. Thurber
    • 2
  • K. Dane Wittrup
    • 1
    • 2
  1. 1.Department of Biological EngineeringMassachusetts Institute of TechnologyCambridgeUSA
  2. 2.Department of Chemical EngineeringMassachusetts Institute of TechnologyCambridgeUSA

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