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Affinity Maturation of an ERBB2-Targeted SPECT Imaging Peptide by In Vivo Phage Display

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Abstract

Purpose

The goal of this study was to improve the pharmacokinetic properties and specificity of an ERBB2-targeted peptide for SPECT imaging.

Procedures

Bacteriophages (phages) displaying the ERBB2 targeting sequence, KCCYSL, flanked by additional random amino acids were used for in vivo selections in mice-bearing ERBB2-expressing MDA-MB-435 human breast xenografts. Phage-displayed peptides were evaluated for ERBB2 and cancer cell binding affinity and specificity in vitro, and one peptide was radiolabeled with 111In-DOTA and biodistribution and SPECT imaging properties were compared to the first generation peptide, 111In-DOTA-KCCYSL.

Results

In vivo phage display selected two peptides, 1-D03 (MEGPSKCCYSLALSH) and 3-G03 (SGTKSKCCYSLRRSS), with higher breast carcinoma cell specificity and similar ErbB2 affinity (236 and 289 nM, respectively) to the first generation peptide. The corresponding radiolabeled probes bound with higher affinity to target cancer cells than 111In-DOTA-KCCYSL; however, only 111In-DOTA-1-D03 demonstrated higher specificity for MDA-MB-435 cells. Biodistribution analysis demonstrated that although 111In-DOTA-1-D03 had slightly reduced tumor uptake (0.661 % ID/g) in comparison to 111In-DOTA-KCCYSL (0.78 %/ID/g), its dramatic improvement in blood clearance led to a significantly higher tumor/blood ratio (6.02:1). Non-specific uptake was also reduced in most organs including heart, lung, muscle, bone, and kidneys. SPECT imaging revealed tumor-specific uptake of 111In-DOTA-1-D03, which was confirmed by blocking with unlabeled 1-D03 peptide.

Conclusions

This is the first evidence that SPECT imaging peptides with improved tumor specificity and pharmacokinetics can be obtained by in vivo phage display affinity maturation. The combination of ERBB2-specific binding, rapid clearance, and tumor specificity may make 1-D03 a viable candidate for clinical imaging studies.

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Acknowledgments

The authors would like to acknowledge the contributions of Jessica Newton-Northup, Marie T. Dickerson, and the VA Biomolecular Imaging Core. Work funded by Department of Veterans Affairs VA Merit 5I0 1BX000964.

Conflict of Interest Statement

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Research Service, Harry S. Truman Veterans Memorial Hospital, Columbia, MO, USA

    Benjamin M. Larimer & Susan L. Deutscher

  2. Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, MO, 65211, USA

    Benjamin M. Larimer & Susan L. Deutscher

  3. Department of Biology, University of Missouri, Columbia, MO, USA

    William D. Thomas & George P. Smith

Authors
  1. Benjamin M. Larimer
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  2. William D. Thomas
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  3. George P. Smith
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  4. Susan L. Deutscher
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Correspondence to Susan L. Deutscher.

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Larimer, B.M., Thomas, W.D., Smith, G.P. et al. Affinity Maturation of an ERBB2-Targeted SPECT Imaging Peptide by In Vivo Phage Display. Mol Imaging Biol 16, 449–458 (2014). https://doi.org/10.1007/s11307-014-0724-5

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  • DOI: https://doi.org/10.1007/s11307-014-0724-5

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