H-type Dimer Formation of Fluorophores: a Mechanism for Activatable, in Vivo Optical Molecular Imaging

Overview

Journal ACS Chem Biol

Publisher American Chemical Society

Specialties Biochemistry
Biology

Date 2009 Jun 2

PMID 19480464

Citations 64

Authors

Mikako Ogawa

Nobuyuki Kosaka

Peter L Choyke

Hisataka Kobayashi

Affiliations

Soon will be listed here.

Abstract

In vivo molecular imaging with target-specific activatable "smart" probes, which yield fluorescence only at the intended target, enables sensitive and specific cancer detection. Dimerization and fluorescence quenching has been shown to occur in concentrated aqueous solutions of various fluorophores. Here, we hypothesized that fluorophore dimerization and quenching after conjugation to targeting proteins can occur at low concentration. This dimerization can be exploited as a mechanism for fluorescence activation. Rhodamine derivatives were conjugated to avidin and trastuzumab, which target D-galactose receptor and HER2/neu antigen, respectively. After conjugation, a large proportion of R6G and TAMRA formed H-type dimers, even at low concentrations, but could be fully dequenched upon dissociation of the dimers to monomers. To demonstrate the fluorescence activation effect during in vivo fluorescence endoscopic molecular imaging, a highly quenched probe, avidin-TAMRA, or a minimally quenched probe, avidin-Alexa488, was administered into mice with ovarian metastases to the peritoneum. The tumors were clearly visualized with avidin-TAMRA, with low background fluorescence; in contrast, the background fluorescence was high for avidin-Alexa488. Thus, H-dimer formation as a mechanism of fluorescence quenching could be used to develop fluorescence activatable probes for in vivo molecular imaging.

Citing Articles

Establishment of a Rapid and Convenient Fluoroimmunoassay Platform Using Antibodies Against PDL1 and HER2.

Choi J, Yun H, Jeong H Curr Issues Mol Biol. 2025; 47(1).

PMID: 39852177 PMC: 11763867. DOI: 10.3390/cimb47010062.

Erythrocyte nano-ghosts with dual optical and magnetic resonance characteristics.

Lee C, Zaman S, Kundra V, Anvari B J Biomed Opt. 2024; 29(8):085001.

PMID: 39165858 PMC: 11333968. DOI: 10.1117/1.JBO.29.8.085001.

Conformational Dynamic Studies of Prokaryotic Potassium Channels Explored by Homo-FRET Methodologies.

Coutinho A, Poveda J, Renart M Methods Mol Biol. 2024; 2796:35-72.

PMID: 38856894 DOI: 10.1007/978-1-0716-3818-7_3.

Fluorophore multimerization on a PEG backbone as a concept for signal amplification and lifetime modulation.

Reiber T, Hubner O, Dose C, Yushchenko D, Resch-Genger U Sci Rep. 2024; 14(1):11882.

PMID: 38789582 PMC: 11126734. DOI: 10.1038/s41598-024-62548-4.

Development of fluorescence-linked immunosorbent assay for rapid detection of Staphylococcus aureus.

Kim J, Yun H, Kim J, Kim W, Lee C, Kim B Appl Microbiol Biotechnol. 2023; 108(1):2.

PMID: 38153552 DOI: 10.1007/s00253-023-12836-2.

References

Fujimoto K, Shimizu H, Inouye M . Unambiguous detection of target DNAs by excimer-monomer switching molecular beacons. J Org Chem. 2004; 69(10):3271-5. DOI: 10.1021/jo049824f. View

Longmire M, Ogawa M, Hama Y, Kosaka N, Regino C, Choyke P . Determination of optimal rhodamine fluorophore for in vivo optical imaging. Bioconjug Chem. 2008; 19(8):1735-42. PMC: 2756081. DOI: 10.1021/bc800140c. View

Hendrickson H, Hendrickson E, Johnson I, Farber S . Intramolecularly quenched BODIPY-labeled phospholipid analogs in phospholipase A(2) and platelet-activating factor acetylhydrolase assays and in vivo fluorescence imaging. Anal Biochem. 1999; 276(1):27-35. DOI: 10.1006/abio.1999.4280. View

Ghose A, Crippen G . Atomic physicochemical parameters for three-dimensional-structure-directed quantitative structure-activity relationships. 2. Modeling dispersive and hydrophobic interactions. J Chem Inf Comput Sci. 1987; 27(1):21-35. DOI: 10.1021/ci00053a005. View

Hama Y, Urano Y, Koyama Y, Kamiya M, Bernardo M, Paik R . A target cell-specific activatable fluorescence probe for in vivo molecular imaging of cancer based on a self-quenched avidin-rhodamine conjugate. Cancer Res. 2007; 67(6):2791-9. DOI: 10.1158/0008-5472.CAN-06-3315. View

Ogawa M, Kosaka N, Longmire M, Urano Y, Choyke P, Kobayashi H . Fluorophore-quencher based activatable targeted optical probes for detecting in vivo cancer metastases. Mol Pharm. 2009; 6(2):386-95. PMC: 2891627. DOI: 10.1021/mp800115t. View

Weissleder R, Pittet M . Imaging in the era of molecular oncology. Nature. 2008; 452(7187):580-9. PMC: 2708079. DOI: 10.1038/nature06917. View

Urano Y, Asanuma D, Hama Y, Koyama Y, Barrett T, Kamiya M . Selective molecular imaging of viable cancer cells with pH-activatable fluorescence probes. Nat Med. 2008; 15(1):104-9. PMC: 2790281. DOI: 10.1038/nm.1854. View

Hama Y, Urano Y, Koyama Y, Gunn A, Choyke P, Kobayashi H . A self-quenched galactosamine-serum albumin-rhodamineX conjugate: a "smart" fluorescent molecular imaging probe synthesized with clinically applicable material for detecting peritoneal ovarian cancer metastases. Clin Cancer Res. 2007; 13(21):6335-43. DOI: 10.1158/1078-0432.CCR-07-1004. View

10.

Bergstrom F, Mikhalyov I, Hagglof P, Wortmann R, Ny T, Johansson L . Dimers of dipyrrometheneboron difluoride (BODIPY) with light spectroscopic applications in chemistry and biology. J Am Chem Soc. 2002; 124(2):196-204. DOI: 10.1021/ja010983f. View

11.

Jiang T, Olson E, Nguyen Q, Roy M, Jennings P, Tsien R . Tumor imaging by means of proteolytic activation of cell-penetrating peptides. Proc Natl Acad Sci U S A. 2004; 101(51):17867-72. PMC: 539314. DOI: 10.1073/pnas.0408191101. View

12.

Kamiya M, Kobayashi H, Hama Y, Koyama Y, Bernardo M, Nagano T . An enzymatically activated fluorescence probe for targeted tumor imaging. J Am Chem Soc. 2007; 129(13):3918-29. PMC: 2555972. DOI: 10.1021/ja067710a. View

13.

Kishimoto H, Kojima T, Watanabe Y, Kagawa S, Fujiwara T, Uno F . In vivo imaging of lymph node metastasis with telomerase-specific replication-selective adenovirus. Nat Med. 2006; 12(10):1213-9. DOI: 10.1038/nm1404. View

14.

Farber S, Pack M, Ho S, Johnson I, Wagner D, Dosch R . Genetic analysis of digestive physiology using fluorescent phospholipid reporters. Science. 2001; 292(5520):1385-8. DOI: 10.1126/science.1060418. View

15.

Hoffman R . The multiple uses of fluorescent proteins to visualize cancer in vivo. Nat Rev Cancer. 2005; 5(10):796-806. DOI: 10.1038/nrc1717. View

16.

Weissleder R, Mahmood U . Molecular imaging. Radiology. 2001; 219(2):316-33. DOI: 10.1148/radiology.219.2.r01ma19316. View

17.

Urano Y . Sensitive and selective tumor imaging with novel and highly activatable fluorescence probes. Anal Sci. 2008; 24(1):51-3. DOI: 10.2116/analsci.24.51. View

18.

Ogawa M, Kosaka N, Choyke P, Kobayashi H . In vivo molecular imaging of cancer with a quenching near-infrared fluorescent probe using conjugates of monoclonal antibodies and indocyanine green. Cancer Res. 2009; 69(4):1268-72. PMC: 2788996. DOI: 10.1158/0008-5472.CAN-08-3116. View

19.

Ogawa M, Regino C, Choyke P, Kobayashi H . In vivo target-specific activatable near-infrared optical labeling of humanized monoclonal antibodies. Mol Cancer Ther. 2009; 8(1):232-9. PMC: 2790275. DOI: 10.1158/1535-7163.MCT-08-0862. View