Production of [F-18]Fluoroannexin for Imaging Apoptosis with PET

John R. Grierson, Kevin J. Yagle, Janet F. Eary, Jonathan F. Tait, Don F. Gibson, Barbara Lewellen, Jeanne Link, Kenneth A. Krohnt

Research output: Contribution to journalArticle

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Abstract

Recombinant human-annexin-V was conjugated with 4-[F-18]fluorobenzoic acid (FBA) via its reaction with the N-hydroxysuccinimidyl ester (FBA-OSu) at pH 8.5. A series of reactions using varying amounts of annexin-V, unlabeled FBA-OSu, and time produced products with different conjugation levels. Products were characterized by mass spectrometry and a cell-binding assay to assess the effect of conjugation. In each case, the conjugated protein was a mixture of proteins with a range of conjugation. Annexin-V could be conjugated with an average of two FBA mole equivalents without decreasing its affinity for red blood cells (Kd 6-10 μM) with exposed phosphatidylserine. An average conjugation of 7.7 (range 3-13) diminished the binding 3-fold. Large-scale production and purification of [F-18]-FBA-OSu from [F-18] fluoride was accomplished within 90 min and in 77% radiochemical yield (decay-corrected to the end of cyclotron bombardment). The conjugation reaction of annexin with [F-18]FBA-OSu was studied with respect to activity level, protein mass, and concentration. Under the most favorable conditions, >25 mCi [F-18]fluoroannexin (FAN) was isolated in 64% yield (decay-corrected for a 22 min conjugation process) from labeling 1.1 mg of annexin-V. A pilot PET imaging study of [F-18]fluoroannexin in normal rats showed high uptake in the renal excretory system and demonstrated sufficient clearance from most other internal organs within 1 h. [F-18]Fluoroannexin should prove useful in imaging targeted apoptosis.

Original languageEnglish (US)
Pages (from-to)373-379
Number of pages7
JournalBioconjugate Chemistry
Volume15
Issue number2
DOIs
StatePublished - Mar 2004
Externally publishedYes

Fingerprint

Cell death
Annexin A5
Apoptosis
Imaging techniques
Acids
Proteins
Cyclotrons
Annexins
Phosphatidylserines
Fluorides
Labeling
Purification
Mass spectrometry
Rats
Assays
Mass Spectrometry
Esters
Blood
Erythrocytes
Cells

ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Grierson, J. R., Yagle, K. J., Eary, J. F., Tait, J. F., Gibson, D. F., Lewellen, B., ... Krohnt, K. A. (2004). Production of [F-18]Fluoroannexin for Imaging Apoptosis with PET. Bioconjugate Chemistry, 15(2), 373-379. https://doi.org/10.1021/bc0300394

Production of [F-18]Fluoroannexin for Imaging Apoptosis with PET. / Grierson, John R.; Yagle, Kevin J.; Eary, Janet F.; Tait, Jonathan F.; Gibson, Don F.; Lewellen, Barbara; Link, Jeanne; Krohnt, Kenneth A.

In: Bioconjugate Chemistry, Vol. 15, No. 2, 03.2004, p. 373-379.

Research output: Contribution to journalArticle

Grierson, JR, Yagle, KJ, Eary, JF, Tait, JF, Gibson, DF, Lewellen, B, Link, J & Krohnt, KA 2004, 'Production of [F-18]Fluoroannexin for Imaging Apoptosis with PET', Bioconjugate Chemistry, vol. 15, no. 2, pp. 373-379. https://doi.org/10.1021/bc0300394
Grierson JR, Yagle KJ, Eary JF, Tait JF, Gibson DF, Lewellen B et al. Production of [F-18]Fluoroannexin for Imaging Apoptosis with PET. Bioconjugate Chemistry. 2004 Mar;15(2):373-379. https://doi.org/10.1021/bc0300394
Grierson, John R. ; Yagle, Kevin J. ; Eary, Janet F. ; Tait, Jonathan F. ; Gibson, Don F. ; Lewellen, Barbara ; Link, Jeanne ; Krohnt, Kenneth A. / Production of [F-18]Fluoroannexin for Imaging Apoptosis with PET. In: Bioconjugate Chemistry. 2004 ; Vol. 15, No. 2. pp. 373-379.
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