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Synthesis of [18F]SiFB: a prosthetic group for direct protein radiolabeling for application in positron emission tomography

Nature Protocols volume 7, pages 1956–1963 (2012)Cite this article

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Abstract

N-Succinimidyl 3-(di-tert-butyl[18F]fluorosilyl)benzoate ([18F]SiFB) is a highly reactive prosthetic group for radiolabeling of proteins for use in positron emission tomography (PET). It is similar to N-succinimidyl-4-[18F]fluorobenzoate ([18F]SFB), the 'gold-standard' prosthetic group for protein 18F-labeling, but can be synthesized using a much shorter and technically easier procedure. A recently reported simple procedure to obtain anhydrous 18F− by avoiding time-consuming azeotropic drying is applied with a slight modification to prevent basic hydrolysis of the active N-hydroxysuccinimide (NHS) ester moiety of [18F]SiFB. The labeling of [18F]SiFB is performed by a fast 18F–19F isotopic exchange (IE) reaction at room temperature (20–25 °C) within 30 min. [18F]SiFB is purified using a C18 cartridge instead of HPLC, further decreasing the overall time required for protein labeling. High specific activities >18.5 GBq μmol−1 (>500 Ci mmol−1) can be obtained. Finally, incubation of [18F]SiFB with the desired protein in an aqueous solution at pH 9, followed by HPLC purification, provides the final solution of the labeled protein ready for in vivo applications.

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Figure 1: 18F-Labeling of proteins using [18F]SiFB ([18F]5).
Figure 2: Radio-TLC of the purified [18F]SiFB.
Figure 3: HPLC chromatogram of [18F]SiFB-labeled RSA ([18F]6) after HPLC purification.

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  • 23 October 2012

     In the version of this article initially published online, Katy Orchowski's name was spelled incorrectly. The surname Orchovski has been corrected to read Orchowski. The error has been corrected in all versions of this article.

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Acknowledgements

We thank the cyclotron crew of the McConnell Brain Imaging Centre for providing fluorine-18. The financial support of the following grant agencies are gratefully acknowledged: Canada Foundation for Innovation project no. 203639 to R.S., Natural Sciences and Engineering Research Council of Canada (NSERC) Strategic Project Grant Program (STPGP) no. 412893 to R.S., Bayern-Quebec-Allianz to S.N., Deutsche Forschungsgemeinschaft (DFG) grant no. WA 2132/3-1 to B.W., and Fonds der Chemischen Industrie to C.W.

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Authors and Affiliations

  1. McConnell Brain Imaging Centre, Montreal Neurological Institute, McGill University, Montreal, Quebec, Canada

    Alexey P Kostikov, Joshua Chin, Katy Orchowski, Esther Schirrmacher & Ralf Schirrmacher

  2. Department of Nuclear Medicine, Hospital of the Ludwig-Maximilians-University, Munich, Germany

    Sabrina Niedermoser & Carmen Wängler

  3. Lehrstuhl für Anorganische Chemie II, Technische Universität Dortmund, Dortmund, Germany

    Klaus Jurkschat & Liuba Iovkova-Berends

  4. Biomedical, Medical Faculty Mannheim, Medical Research Center Mannheim, Heidelberg University, Mannheim, Germany

    Carmen Wängler

  5. Department of Clinical Radiology and Nuclear Medicine, Molecular Imaging and Radiochemistry, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany

    Björn Wängler

Authors
  1. Alexey P Kostikov
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  2. Joshua Chin
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  3. Katy Orchowski
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  4. Esther Schirrmacher
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  6. Klaus Jurkschat
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  7. Liuba Iovkova-Berends
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  8. Carmen Wängler
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  9. Björn Wängler
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Contributions

A.P.K., S.N., J.C., K.O., E.S., L.I.-B., K.J. and C.W. performed the experimental work. R.S., C.W. and B.W. wrote the manuscript.

Corresponding author

Correspondence to Ralf Schirrmacher.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Figure 1

1H NMR spectrum of compound 1 (PDF 426 kb)

Supplementary Figure 2

13C NMR spectrum of compound 1 (PDF 445 kb)

Supplementary Figure 3

1H NMR spectrum of compound 2 (PDF 441 kb)

Supplementary Figure 4

13C NMR spectrum of compound 2 (PDF 435 kb)

Supplementary Figure 5

1H NMR spectrum of compound 3 (PDF 429 kb)

Supplementary Figure 6

13C NMR spectrum of compound 3 (PDF 428 kb)

Supplementary Figure 7

1H NMR spectrum of compound 4 (PDF 428 kb)

Supplementary Figure 8

13C NMR spectrum of compound 4 (PDF 429 kb)

Supplementary Figure 9

1H NMR spectrum of compound 5 (PDF 431 kb)

Supplementary Figure 10

13C NMR spectrum of compound 5 (PDF 439 kb)

Supplementary Figure 11

Radio-TLC of [18F]5 (PDF 485 kb)

Supplementary Figure 12

HPLC chromatogram of [18F]5-RSA conjugate (PDF 87 kb)

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Kostikov, A., Chin, J., Orchowski, K. et al. Synthesis of [18F]SiFB: a prosthetic group for direct protein radiolabeling for application in positron emission tomography. Nat Protoc 7, 1956–1963 (2012). https://doi.org/10.1038/nprot.2012.110

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