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FN3 Protein Conjugates for Cancer Diagnosis and Imaging Studies

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Bioconjugation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2033))

Abstract

Bioconjugation of biologically useful proteins is in great demand (e.g., conjugation to biotins, metal chelators, and drug carriers to target specific tissues for both in vitro and in vivo use). These conjugates provide widespread opportunities for various biological and biomedical applications. Evolving state-of-the-art protein conjugation strategies have led to the development of many affinity ligands, including for cancer imaging and diagnosis. However, to achieve the desirable protein conjugates, there are many challenges that remain to be addressed in order to obtain a reproducible procedure for all proteins and ligands. These include a control over the protein modification and the efficiency of the conjugation while retaining the original biological protein affinity postmodification. Here we present detailed conjugation methods for the human fibronectin tenth type III domain (FN3) protein scaffold for use in preclinical PET imaging. More specifically, this chapter provides detailed methods to produce a FN3 and a FN3-chelator-conjugate, its labeling with the radionuclide 64-Cu, and its use for noninvasive PET imaging in mice.

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Acknowledgments

We would like to acknowledge the SCi3 Core imaging facilities and the Canary Center at Stanford for providing technical support.

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Authors and Affiliations

  1. Department of Radiology, James H. Clark Center, Stanford University, Stanford, CA, USA

    Arutselvan Natarajan & Lotfi Abou-Elkacem

Authors
  1. Arutselvan Natarajan
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  2. Lotfi Abou-Elkacem
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Corresponding author

Correspondence to Arutselvan Natarajan .

Editor information

Editors and Affiliations

  1. Cellular and Molecular Immunology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium

    Sam Massa

  2. In Vivo Cellular and Molecular Imaging Laboratory (ICMI), Vrije Universiteit Brussel, Brussels, Belgium

    Nick Devoogdt

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Natarajan, A., Abou-Elkacem, L. (2019). FN3 Protein Conjugates for Cancer Diagnosis and Imaging Studies. In: Massa, S., Devoogdt, N. (eds) Bioconjugation. Methods in Molecular Biology, vol 2033. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9654-4_20

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  • DOI: https://doi.org/10.1007/978-1-4939-9654-4_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9653-7

  • Online ISBN: 978-1-4939-9654-4

  • eBook Packages: Springer Protocols

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