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Streptavidin in Antibody Pretargeting. 2. Evaluation of Methods for Decreasing Localization of Streptavidin to Kidney while Retaining Its Tumor Binding Capacity

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    Streptavidin in Antibody Pretargeting. 2. Evaluation of Methods for Decreasing Localization of Streptavidin to Kidney while Retaining Its Tumor Binding Capacity
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    Departments of Radiation Oncology, Urology, and Bioengineering, University of Washington, Seattle, Washington 98195
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    Bioconjugate Chemistry

    Cite this: Bioconjugate Chem. 1998, 9, 3, 322–330
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    https://doi.org/10.1021/bc970182v
    Published April 4, 1998
    Copyright © 1998 American Chemical Society
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    Abstract

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    An investigation has been conducted to determine if the kidney localization of recombinant streptavidin can be decreased to improve its characteristics in pretargeting protocols. Three different methods of accomplishing this were evaluated. The first method, blocking kidney uptake with a preadministration of recombinant streptavidin in which biotin occupied all of the binding sites, was unsuccessful. In a second method, l-lysine administration was used to block kidney localization. This method worked well, decreasing the concentration to 29% of the unmodified amount at 8 h postinjection. However, this method suffered from a requirement for constant infusion of lysine during the period of observation. A third method, use of succinylated recombinant streptavidin, was found to be the best approach. Succinylation of streptavidin was readily accomplished with very good protein recovery. With the succinylated streptavidin, the kidney concentration was only 14% of that of nonmodified streptavidin at 4 h postinjection. While these results demonstrated that the concentration of streptavidin could be decreased in the kidney, it was important to assess whether the tumor colocalization of streptavidin with biotinylated antibody was affected under those conditions. As part of our continuing investigation of pretargeting, a new water-solubilized biotinidase-stabilized biotinylation reagent was prepared. Using that reagent in a pretargeting experiment, an equivalent quantity of succinylated recombinant streptavidin as biotinylated antibody Fab‘ was localized in a tumor xenograft model. In that experiment, the kidney concentration was decreased to less than 10% of that obtained with unmodified recombinant streptavidin at 24 h postinjection. The results of our investigation have demonstrated that succinylation of streptavidin improves its distribution characteristics for pretargeting applications. The fact that succinylated streptavidin has no specific tissue localization should allow its use as a carrier of radioactivity in “two-step” pretargeting protocols.

    Copyright © 1998 American Chemical Society

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     Address correspondence to D. Scott Wilbur, Ph.D., Department of Radiation Oncology, University of Washington, 2121 N. 35th St., Seattle, WA 98103-9103. Phone:  206-685-3085. Fax:  206-685-9630. E-mail:  [email protected].

    †

     Department of Radiation Oncology.

    ‡

     Department of Urology.

    §

     Department of Bioengineering.

    Supporting Information Available

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    HPLC chromatograms and 1H NMR spectra of the new compounds (5 and 7) prepared in this paper (3 pages). Ordering information is given on any current masthead page.

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    Bioconjugate Chemistry

    Cite this: Bioconjugate Chem. 1998, 9, 3, 322–330
    Click to copy citationCitation copied!
    https://doi.org/10.1021/bc970182v
    Published April 4, 1998
    Copyright © 1998 American Chemical Society
    Request reuse permissions

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