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Quantitating Antibody Uptake In Vivo: Conditional Dependence on Antigen Expression Levels

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Abstract

Purpose

Antibodies form an important class of cancer therapeutics, and there is intense interest in using them for imaging applications in diagnosis and monitoring of cancer treatment. Despite the expanding body of knowledge describing pharmacokinetic and pharmacodynamic interactions of antibodies in vivo, discrepancies remain over the effect of antigen expression level on tumoral uptake with some reports indicating a relationship between uptake and expression and others showing no correlation.

Procedures

Using a cell line with high epithelial cell adhesion molecule expression and moderate epidermal growth factor receptor expression, fluorescent antibodies with similar plasma clearance were imaged in vivo. A mathematical model and mouse xenograft experiments were used to describe the effect of antigen expression on uptake of these high-affinity antibodies.

Results

As predicted by the theoretical model, under subsaturating conditions, uptake of the antibodies in such tumors is similar because localization of both probes is limited by delivery from the vasculature. In a separate experiment, when the tumor is saturated, the uptake becomes dependent on the number of available binding sites. In addition, targeting of small micrometastases is shown to be higher than larger vascularized tumors.

Conclusions

These results are consistent with the prediction that high affinity antibody uptake is dependent on antigen expression levels for saturating doses and delivery for subsaturating doses. It is imperative for any probe to understand whether quantitative uptake is a measure of biomarker expression or transport to the region of interest. The data provide support for a predictive theoretical model of antibody uptake, enabling it to be used as a starting point for the design of more efficacious therapies and timely quantitative imaging probes.

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Abbreviations

EGFR:

Epidermal Growth Factor Receptor

EpCAM:

Epithelial Cell Adhesion Molecule

VT680:

VivoTag 680 fluorescent dye

AF750:

AlexaFluor 750 fluorescent dye

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Acknowledgments

This work was supported by grants P50 CA86355, U24 CA092782, and T32 CA079443.

Conflict of Interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA

    Greg M. Thurber & Ralph Weissleder

  2. Simches Research Building, 185 Cambridge St., Boston, MA, 02114, USA

    Ralph Weissleder

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  1. Greg M. Thurber
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  2. Ralph Weissleder
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Correspondence to Ralph Weissleder.

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Thurber, G.M., Weissleder, R. Quantitating Antibody Uptake In Vivo: Conditional Dependence on Antigen Expression Levels. Mol Imaging Biol 13, 623–632 (2011). https://doi.org/10.1007/s11307-010-0397-7

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  • DOI: https://doi.org/10.1007/s11307-010-0397-7

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