Abstract
Purpose
Although a preparation method for F-18-labeled proteins that used a cell-free translation system and 4-[18F]fluoro-L-proline instead of L-proline has been reported, its introduction depends on amino acid sequences of target proteins. The purpose of the study was to propose site-specific labeling method of F-18 by using cell-free translation systems supplemented with an engineered orthogonal aminoacyl-tRNA synthetase derived from Methanocaldococcus jannaschii (pCNF-RS)/suppressor tRNA (tRNACUAopt) pair, O-2-[18F]fluoroethyl-L-tyrosine ([18F]FET), and template DNA inserted with an amber codon.
Procedures
[18F]FET was prepared from the corresponding precursor and determined whether [18F]FET could be incorporated into an affibody molecule for human epidermal growth factor receptor type 2 (HER2; ZHER2:342) as the 21st amino acid used with the pCNF-RS-tRNACUAopt pair and template DNA inserted with an amber codon in a cell-free translation system. Using SKOV-3 cells, we performed an in vitro binding assay of [18F]FET-ZHER2:342. Furthermore, in vivo positron emission tomography (PET) imaging in SKOV-3 xenograft-bearing mice was performed after the intravenous administration of [18F]FET-ZHER2:342.
Results
[18F]FET was successfully incorporated into proteins by using commercially available cell-free protein synthesis reagents with a pCNF-RS-tRNACUAopt pair and template DNA of the desired proteins inserted with an amber codon. The mean radiochemical yield (non-decay-corrected) of [18F]FET-ZHER2:342 was 6.5 ± 4.1 %. An in vitro cell binding assay revealed that SKOV-3 cells–bound [18F]FET-ZHER2:342 expressed HER2. The in vivo PET imaging in SKOV-3 xenograft-bearing mice revealed that [18F]FET-ZHER2:342 accumulated in SKOV-3 xenografts.
Conclusion
The method proposed in this study might be useful for preparing proteins with F-18 and molecular imaging in the preclinical development.
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References
Koerber JT, Thomsen ND, Hannigan BT, Degrado WF, Wells JA (2013) Nature-inspired design of motif-specific antibody scaffolds. Nat Biotechnol 31:916–921
Colombo I, Overchuk M, Chen J, Reilly RM, Zheng G, Lheureux S (2017) Molecular imaging in drug development: update and challenges for radiolabeled antibodies and nanotechnology. Methods 130:23–35
Moek KL, Giesen D, Kok IC, de Groot DJA, Jalving M, Fehrmann RSN, Lub-de Hooge MN, Brouwers AH, de Vries EGE (2017) Theranostics using antibodies and antibody-related therapeutics. J Nucl Med 58:83S–90S
Tolmachev V, Orlova A, Nilsson FY, Feldwisch J, Wennborg A, Abrahmsén L (2007) Affibody molecules: potential for in vivo imaging of molecular targets for cancer therapy. Expert Opin Biol Ther 7:555–568
Orlova A, Magnusson M, Eriksson TL et al (2006) Tumor imaging using a picomolar affinity HER2 binding affibody molecule. Cancer Res 66:4339–4348
Wallberg H, Grafstrom J, Cheng Q, Lu L, Martinsson Ahlzen HS, Samen E, Thorell JO, Johansson K, Dunas F, Olofsson MH, Stone-Elander S, Arner ESJ, Stahl S (2012) HER2-positive tumors imaged within 1 hour using a site-specifically 11C-labeled Sel-tagged affibody molecule. J Nucl Med 53:1446–1453
Glaser M, Iveson P, Hoppmann S, Indrevoll B, Wilson A, Arukwe J, Danikas A, Bhalla R, Hiscock D (2013) Three methods for 18F labeling of the HER2-binding affibody molecule ZHER2:2891 including preclinical assessment. J Nucl Med 54:1981–1988
Harada R, Furumoto S, Yoshikawa T, Ishikawa Y, Shibuya K, Okamura N, Ishiwata K, Iwata R, Yanai K (2016) Synthesis and characterization of 18F-interleukin-8 using a cell-free translation system and 4-18F-fluoro-L-proline. J Nucl Med 57:634–639
Harada R, Furumoto S, Yoshikawa T, Ishikawa Y, Shibuya K, Okamura N, Iwata R, Yanai K (2012) Synthesis of [11C]interleukin 8 using a cell-free translation system and L-[11C]methionine. Nucl Med Biol 39:155–160
Shimizu Y, Inoue A, Tomari Y, Suzuki T, Yokogawa T, Nishikawa K, Ueda T (2001) Cell-free translation reconstituted with purified components. Nat Biotechnol 19:751–755
Liu CC, Schultz PG (2010) Adding new chemistries to the genetic code. Annu Rev Biochem 79:413–444
Wang L, Brock A, Herberich B, Schultz PG (2001) Expanding the genetic code of Escherichia coli. Science 292:498–500
Goerke AR, Swartz JR (2008) Development of cell-free protein synthesis platforms for disulfide bonded proteins. Biotechnol Bioeng 99:351–367
Ozawa K, Loscha KV, Kuppan KV, Loh CT, Dixon NE, Otting G (2012) High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites. Biochem Biophys Res Commun 418:652–656
Young DD, Young TS, Jahnz M, Ahmad I, Spraggon G, Schultz PG (2011) An evolved aminoacyl-tRNA synthetase with atypical polysubstrate specificity. Biochemistry 50:1894–1900
Tolmachev V, Hofstrom C, Malmberg J et al (2010) HEHEHE-tagged affibody molecule may be purified by IMAC, is conveniently labeled with [99mTc(CO3)]+, and shows improved biodistribution with reduced hepatic radioactivity accumulation. Bioconjug Chem 21:2013–2022
Young TS, Ahmad I, Yin JA, Schultz PG (2010) An enhanced system for unnatural amino acid mutagenesis in E. coli. J Mol Biol 395:361–374
Iwata R, Pascali C, Terasaki K, Ishikawa Y, Furumoto S, Yanai K (2018) Practical microscale one-pot radiosynthesis of (18) F-labeled probes. J Labelled Comp Radiopharm 61:540–549
Hamacher K, Coenen HH (2002) Efficient routine production of the 18F-labelled amino acid O-2-18F fluoroethyl-L-tyrosine. Appl Radiat Isot 57:853–856
Loening AM, Gambhir SS (2003) AMIDE: a free software tool for multimodality medical image analysis. Mol Imaging 2:131–137
Qi Y, Liu X, Li J, Yao H, Yuan S (2017) Fluorine-18 labeled amino acids for tumor PET/CT imaging. Oncotarget 8:60581–60588
Rapp M, Heinzel A, Galldiks N, Stoffels G, Felsberg J, Ewelt C, Sabel M, Steiger HJ, Reifenberger G, Beez T, Coenen HH, Floeth FW, Langen KJ (2013) Diagnostic performance of 18F-FET PET in newly diagnosed cerebral lesions suggestive of glioma. J Nucl Med 54:229–235
Dunet V, Rossier C, Buck A, Stupp R, Prior JO (2012) Performance of 18F-fluoro-ethyl-tyrosine (18F-FET) PET for the differential diagnosis of primary brain tumor: a systematic review and Metaanalysis. J Nucl Med 53:207–214
Kramer-Marek G, Kiesewetter DO, Martiniova L, Jagoda E, Lee SB, Capala J (2008) [18F]FBEM-Z(HER2:342)-affibody molecule-a new molecular tracer for in vivo monitoring of HER2 expression by positron emission tomography. Eur J Nucl Med Mol Imaging 35:1008–1018
Morris O, Fairclough M, Grigg J, et al. (2018) A review of approaches to (18) F radiolabelling affinity peptides and proteins. J Labelled Comp Radiopharm
Eigenbrot C, Ultsch M, Dubnovitsky A, Abrahmsen L, Hard T (2010) Structural basis for high-affinity HER2 receptor binding by an engineered protein. Proc Natl Acad Sci U S A 107:15039–15044
Wester HJ, Herz M, Weber W, Heiss P, Senekowitsch-Schmidtke R, Schwaiger M, Stöcklin G (1999) Synthesis and radiopharmacology of O-(2-[18F]fluoroethyl)-L-tyrosine for tumor imaging. J Nucl Med 40:205–212
Stahl S, Graslund T, Eriksson Karlstrom A et al (2017) Affibody molecules in biotechnological and medical applications. Trends Biotechnol 35:691–712
Ahlgren S, Orlova A, Wallberg H, Hansson M, Sandstrom M, Lewsley R, Wennborg A, Abrahmsen L, Tolmachev V, Feldwisch J (2010) Targeting of HER2-expressing tumors using 111In-ABY-025, a second-generation affibody molecule with a fundamentally reengineered scaffold. J Nucl Med 51:1131–1138
Orlova A, Tolmachev V, Pehrson R, Lindborg M, Tran T, Sandstrom M, Nilsson FY, Wennborg A, Abrahmsen L, Feldwisch J (2007) Synthetic affibody molecules: a novel class of affinity ligands for molecular imaging of HER2-expressing malignant tumors. Cancer Res 67:2178–2186
Acknowledgments
This study was supported by Grants-in-Aid for Scientific Research (16 K15314) from the Japan Society for the Promotion of Science and the Ministry of Education, Culture, Sports, Science, and Technology, Japan. We wish to acknowledge the contribution of Mr. Takahiro Morito for technical support and the staff at the Cyclotron and Radioisotope Center of Tohoku University for the HM-12 cyclotron operation. We acknowledge the support of the Biomedical Research Core of Tohoku University Graduate School of Medicine.
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Yanai, A., Harada, R., Iwata, R. et al. Site-Specific Labeling of F-18 Proteins Using a Supplemented Cell-Free Protein Synthesis System and O-2-[18F]Fluoroethyl-L-Tyrosine: [18F]FET-HER2 Affibody Molecule. Mol Imaging Biol 21, 529–537 (2019). https://doi.org/10.1007/s11307-018-1266-z
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DOI: https://doi.org/10.1007/s11307-018-1266-z