NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.

Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2013.

Cover of Molecular Imaging and Contrast Agent Database (MICAD)

Molecular Imaging and Contrast Agent Database (MICAD) [Internet].

Show details

Quantum dot-trastuzumab

QT
, PhD
National Center for Biotechnology Information, NLM, NIH, Bethesda, MD
Corresponding author.

Created: ; Last Update: February 24, 2011.

Chemical name:Quantum dot-trastuzumab
Abbreviated name:QT, QD-T
Synonym:Quantum dot-Herceptin®
Agent category:Antibody
Target:Epidermal growth factor receptor (EGFR), HER2
Target category:Receptor
Method of detection:Optical, near-infrared (NIR) fluorescence
Source of signal:Quantum dot (QD)
Activation:No
Studies:
  • Checkbox In vitro
  • Checkbox Rodents

Click on protein, nucleotide (RefSeq), and gene for more information about HER2.

Background

[PubMed]

Epidermal growth factor (EGF) is a 53-amino acid cytokine (6.2 kDa) secreted by ectodermic cells, monocytes, kidneys and duodenal glands (1). EGF stimulates growth of epidermal and epithelial cells. EGF with at least seven other growth factors and their transmembrane receptor kinases play important roles in cell proliferation, survival, adhesion, migration and differentiation. The EGF receptor (EGFR) family consists of four transmembrane receptors, including EGFR (HER1/erbB-1), HER2 (erbB-2/neu), HER3 (erbB-3) and HER4 (erbB-4) (2). HER1, HER3 and HER4 comprise three major functional domains: an extracellular ligand-binding domain, a hydrophobic transmembrane domain and a cytoplasmic tyrosine kinase domain. No ligand has been clearly identified for HER2. However, HER2 can be activated as a result of ligand binding to other HER receptors with the formation of receptor homodimers and/or heterodimers (3). HER1 as well as HER2 are overexpressed on many solid tumor cells such as breast, non-small-cell lung, head and neck, and colon cancer (4-6). The high levels of HER1 and HER2 expression on cancer cells are associated with a poor prognosis (7-10).

Optical fluorescence imaging is increasingly used to monitor biological functions of specific targets in small animals (11-13). However, the intrinsic fluorescence of biomolecules poses a problem when fluorophores that absorb visible light (350-700 nm) are used. Near-infrared (NIR) fluorescence (700-1,000 nm) detection avoids the background fluorescence interference of natural biomolecules, providing a high contrast between target and background tissues. NIR fluorophores have a wider dynamic range and minimal background as a result of reduced scattering compared with visible fluorescence detection. They also have high sensitivity, resulting from low fluorescence background, and high extinction coefficients, which provide high quantum yields. The NIR region is also compatible with solid-state optical components, such as diode lasers and silicon detectors. NIR fluorescence imaging is becoming a non-invasive alternative to radionuclide imaging in small animals (14, 15).

Fluorescent semiconductor quantum dots (QDs) are nanocrystals made of CdSe/CdTe-ZnS with radii of 1-10 nm (16-18). They can be tuned to emit in a range of wavelengths by changing their sizes and composition, thus providing broad excitation profiles and high absorption coefficients. They have narrow and symmetric emission spectra with long, excited-state lifetimes, 20-50 ns, as compared with 1-10 ns of fluorescent dyes. They process good quantum yields of 40-90% and high extinction coefficients. They are more photo-stable than conventional organic dyes. They can be coated and capped with hydrophilic materials for additional conjugations with biomolecules, such as peptides, antibodies, nucleic acids, and small organic compounds, which were tested in vitro and in vivo (18-22). Although many cells have been labeled with QDs in vitro with little cytotoxicity, there are only limited studies of long-term toxicity of QDs in small animals (23-31). However, little is known about the toxicity and the mechanisms of clearance and metabolism of QDs in humans.

Trastuzumab is a humanized IgG1 monoclonal antibody (mAb) against the extracellular domain of recombinant HER2 with an affinity constant (Kd) of 0.1 nM (32). Cardiotoxicity is the most serious complication of using trastuzumab in humans with breast cancer (33). One potential application of a radiolabeled anti-HER2 MAb is the pretreatment imaging of breast cancer patients to predict the therapeutic efficacy of trastuzumab. 111In-Trastuzumab, Cy5.5-trastuzumab, and 68Ga-trastuzumab -F(ab')2 have been developed for imaging of human breast cancer (34-38). Trastuzumab has also been successfully coupled with quantum dots for optical imaging of HER2 in tumors in mice (39).

Synthesis

[PubMed]

Commercially available QD-antibody labeling kit was used to conjugate trastuzumab with QDs coated with polyethylene glycol to form QD-trastuzumab (QT) nanoparticles (39). In brief, QDs were first activated with the heterobifunctional cross-linker 4-(maleimidomethyl)-1-cyclohexanecarboxylic acid N-hydroxysuccinimide ester (SMCC) to yield a maleimide-nanocrystal surface. Excess SMCC was removed by column chromatography. Trastuzumab was then reduced and fragmented by dithiothreitol (DTT) to expose free sulfhydryl groups, and excess DTT was removed by column chromatography. The activated QDs were covalently coupled with reduced antibody fragments and the reaction was quenched with ß-mercaptoethanol. QT nanoparticles were purified by gel-filtration chromatography. The molar ratio of trastuzumab fragments to QD was estimated to be ~3.0 by spectrometric analyses. The hydrodynamic diameter of QT was not reported.

In Vitro Studies: Testing in Cells and Tissues

[PubMed]

Tada et al. (39) performed cell-binding assays with QT nanoparticles using human KPL-4 breast cancer cells (overexpressing HER2) and MDA-MB-231 cancer cells (low HER2 expression). Incubation of 10 nM QT nanoparticles for 30 min at 37°C showed that higher fluorescence intensity (a.u.) was observed in the KPL-4 cells (7,000 a.u.) than the MDA-MB-231 cells (<1,000 a.u.). Incubation of 30-100 nM QT nanoparticles provided even higher fluorescence intensity in the KPL-4 cells. On the other hand, binding of control QDs in both cell types was low (< 500 a.u.). Similar results were obtained using fluorescence flow cytometry. Pretreatment of KPL-4 cells with 100 nM trastuzumab before incubation with 100 nM QT nanoparticles showed little fluorescence could be detected on the KPL-4 cells.

Animal Studies

Rodents

[PubMed]

Tada et al. (39) studied the accumulation of QT nanoparticles in nude mice bearing KPL-4 tumors using a fluorescence detection system through a dorsal skinfold chamber after injection of QT nanoparticles (200 nmol/mouse). Movement of single QT nanoparticles was imaged at a video rate of 33 ms/frame with a resolution of 30 nm. The speed of movement was calculated from positional changes of the centroid of the QT images with time. The membranes of the KPL-4 cancer cells were clearly delineated with single QT nanoparticle at 6 h after injection. QT nanoparticles were internalized into the perinuclear region of the cancer cells at 24 h after injection. Six stages of movement of QT nanoparticles were detected: (a) blood vessel circulation (100-600 μm/s), (b) extravasation (1-4 μm/s), (c) diffusion movement into the extracellular space (0.0014 μm2/s), (d) binding to HER2 on the cell membrane (200-400 nm/s), (e) movement from the cell membrane to the perinuclear region after endocytosis (100-300 nm/s), and (f) movement to nuclear region (~600 nm/s).

Other Non-Primate Mammals

[PubMed]

No publication is currently available.

Non-Human Primates

[PubMed]

No publication is currently available.

Human Studies

[PubMed]

No publication is currently available.

References

1.
Carpenter G., Cohen S. Epidermal growth factor. J Biol Chem. 1990;265(14):7709–12. [PubMed: 2186024]
2.
Yarden Y. The EGFR family and its ligands in human cancer. signalling mechanisms and therapeutic opportunities. Eur J Cancer. 2001;37 Suppl 4:S3–8. [PubMed: 11597398]
3.
Rubin I., Yarden Y. The basic biology of HER2. Ann Oncol. 2001;12 Suppl 1:S3–8. [PubMed: 11521719]
4.
Grunwald V., Hidalgo M. Developing inhibitors of the epidermal growth factor receptor for cancer treatment. J Natl Cancer Inst. 2003;95(12):851–67. [PubMed: 12813169]
5.
Mendelsohn J. Anti-epidermal growth factor receptor monoclonal antibodies as potential anti-cancer agents. J Steroid Biochem Mol Biol. 1990;37(6):889–92. [PubMed: 2285602]
6.
Yasui W., Sumiyoshi H., Hata J., Kameda T., Ochiai A., Ito H., Tahara E. Expression of epidermal growth factor receptor in human gastric and colonic carcinomas. Cancer Res. 1988;48(1):137–41. [PubMed: 2446740]
7.
Ang K.K., Berkey B.A., Tu X., Zhang H.Z., Katz R., Hammond E.H., Fu K.K., Milas L. Impact of epidermal growth factor receptor expression on survival and pattern of relapse in patients with advanced head and neck carcinoma. Cancer Res. 2002;62(24):7350–6. [PubMed: 12499279]
8.
Costa S., Stamm H., Almendral A., Ludwig H., Wyss R., Fabbro D., Ernst A., Takahashi A., Eppenberger U. Predictive value of EGF receptor in breast cancer. Lancet. 1988;2(8622):1258. [PubMed: 2903994]
9.
Ethier S.P. Growth factor synthesis and human breast cancer progression. J Natl Cancer Inst. 1995;87(13):964–73. [PubMed: 7629883]
10.
Yarden Y. Biology of HER2 and its importance in breast cancer. Oncology. 2001;61 Suppl 2:1–13. [PubMed: 11694782]
11.
Achilefu S. Lighting up tumors with receptor-specific optical molecular probes. Technol Cancer Res Treat. 2004;3(4):393–409. [PubMed: 15270591]
12.
Ntziachristos V., Bremer C., Weissleder R. Fluorescence imaging with near-infrared light: new technological advances that enable in vivo molecular imaging. Eur Radiol. 2003;13(1):195–208. [PubMed: 12541130]
13.
Becker A., Hessenius C., Licha K., Ebert B., Sukowski U., Semmler W., Wiedenmann B., Grotzinger C. Receptor-targeted optical imaging of tumors with near-infrared fluorescent ligands. Nat Biotechnol. 2001;19(4):327–31. [PubMed: 11283589]
14.
Robeson W., Dhawan V., Belakhlef A., Ma Y., Pillai V., Chaly T., Margouleff C., Bjelke D., Eidelberg D. Dosimetry of the dopamine transporter radioligand 18F-FPCIT in human subjects. J Nucl Med. 2003;44(6):961–6. [PubMed: 12791826]
15.
Tung C.H. Fluorescent peptide probes for in vivo diagnostic imaging. Biopolymers. 2004;76(5):391–403. [PubMed: 15389488]
16.
Zheng G., Li H., Yang K., Blessington D., Licha K., Lund-Katz S., Chance B., Glickson J.D. Tricarbocyanine cholesteryl laurates labeled LDL: new near infrared fluorescent probes (NIRFs) for monitoring tumors and gene therapy of familial hypercholesterolemia. Bioorg Med Chem Lett. 2002;12(11):1485–8. [PubMed: 12031325]
17.
Gao X., Nie S. Quantum dot-encoded beads. Methods Mol Biol. 2005;303:61–71. [PubMed: 15923675]
18.
Michalet X., Pinaud F.F., Bentolila L.A., Tsay J.M., Doose S., Li J.J., Sundaresan G., Wu A.M., Gambhir S.S., Weiss S. Quantum dots for live cells, in vivo imaging, and diagnostics. Science. 2005;307(5709):538–44. [PMC free article: PMC1201471] [PubMed: 15681376]
19.
Alivisatos A.P., Gu W., Larabell C. Quantum dots as cellular probes. Annu Rev Biomed Eng. 2005;7:55–76. [PubMed: 16004566]
20.
Hilger I., Leistner Y., Berndt A., Fritsche C., Haas K.M., Kosmehl H., Kaiser W.A. Near-infrared fluorescence imaging of HER-2 protein over-expression in tumour cells. Eur Radiol. 2004;14(6):1124–9. [PubMed: 15118831]
21.
Medintz I.L., Uyeda H.T., Goldman E.R., Mattoussi H. Quantum dot bioconjugates for imaging, labelling and sensing. Nat Mater. 2005;4(6):435–46. [PubMed: 15928695]
22.
Smith A.M., Gao X., Nie S. Quantum dot nanocrystals for in vivo molecular and cellular imaging. Photochem Photobiol. 2004;80(3):377–85. [PubMed: 15623319]
23.
Akerman M.E., Chan W.C., Laakkonen P., Bhatia S.N., Ruoslahti E. Nanocrystal targeting in vivo. Proc Natl Acad Sci U S A. 2002;99(20):12617–21. [PMC free article: PMC130509] [PubMed: 12235356]
24.
Braydich-Stolle L., Hussain S., Schlager J.J., Hofmann M.C. In vitro cytotoxicity of nanoparticles in mammalian germline stem cells. Toxicol Sci. 2005;88(2):412–9. [PMC free article: PMC2911231] [PubMed: 16014736]
25.
Kim S., Lim Y.T., Soltesz E.G., De Grand A.M., Lee J., Nakayama A., Parker J.A., Mihaljevic T., Laurence R.G., Dor D.M., Cohn L.H., Bawendi M.G., Frangioni J.V. Near-infrared fluorescent type II quantum dots for sentinel lymph node mapping. Nat Biotechnol. 2004;22(1):93–7. [PMC free article: PMC2346610] [PubMed: 14661026]
26.
Gao X., Cui Y., Levenson R.M., Chung L.W., Nie S. In vivo cancer targeting and imaging with semiconductor quantum dots. Nat Biotechnol. 2004;22(8):969–76. [PubMed: 15258594]
27.
Han M., Gao X., Su J.Z., Nie S. Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules. Nat Biotechnol. 2001;19(7):631–5. [PubMed: 11433273]
28.
Lovric J., Bazzi H.S., Cuie Y., Fortin G.R., Winnik F.M., Maysinger D. Differences in subcellular distribution and toxicity of green and red emitting CdTe quantum dots. J Mol Med. 2005;83(5):377–85. [PubMed: 15688234]
29.
Ohnishi S., Lomnes S.J., Laurence R.G., Gogbashian A., Mariani G., Frangioni J.V. Organic alternatives to quantum dots for intraoperative near-infrared fluorescent sentinel lymph node mapping. Mol Imaging. 2005;4(3):172–81. [PubMed: 16194449]
30.
Shiohara A., Hoshino A., Hanaki K., Suzuki K., Yamamoto K. On the cyto-toxicity caused by quantum dots. Microbiol Immunol. 2004;48(9):669–75. [PubMed: 15383704]
31.
Soltesz E.G., Kim S., Laurence R.G., DeGrand A.M., Parungo C.P., Dor D.M., Cohn L.H., Bawendi M.G., Frangioni J.V., Mihaljevic T. Intraoperative sentinel lymph node mapping of the lung using near-infrared fluorescent quantum dots. Ann Thorac Surg. 2005;79(1):269–77. [PMC free article: PMC1421510] [PubMed: 15620956]
32.
Carter P., Presta L., Gorman C.M., Ridgway J.B., Henner D., Wong W.L., Rowland A.M., Kotts C., Carver M.E., Shepard H.M. Humanization of an anti-p185HER2 antibody for human cancer therapy. Proc Natl Acad Sci U S A. 1992;89(10):4285–9. [PMC free article: PMC49066] [PubMed: 1350088]
33.
Goldenberg M.M. Trastuzumab, a recombinant DNA-derived humanized monoclonal antibody, a novel agent for the treatment of metastatic breast cancer. Clin Ther. 1999;21(2):309–18. [PubMed: 10211534]
34.
Perik P.J., Lub-De Hooge M.N., Gietema J.A., van der Graaf W.T., de Korte M.A., Jonkman S., Kosterink J.G., van Veldhuisen D.J., Sleijfer D.T., Jager P.L., de Vries E.G. Indium-111-labeled trastuzumab scintigraphy in patients with human epidermal growth factor receptor 2-positive metastatic breast cancer. J Clin Oncol. 2006;24(15):2276–82. [PubMed: 16710024]
35.
Lub-de Hooge M.N., Kosterink J.G., Perik P.J., Nijnuis H., Tran L., Bart J., Suurmeijer A.J., de Jong S., Jager P.L., de Vries E.G. Preclinical characterisation of 111In-DTPA-trastuzumab. Br J Pharmacol. 2004;143(1):99–106. [PMC free article: PMC1575276] [PubMed: 15289297]
36.
Garmestani K., Milenic D.E., Plascjak P.S., Brechbiel M.W. A new and convenient method for purification of 86Y using a Sr(II) selective resin and comparison of biodistribution of 86Y and 111In labeled Herceptin. Nucl Med Biol. 2002;29(5):599–606. [PubMed: 12088731]
37.
Smith-Jones P.M., Solit D., Afroze F., Rosen N., Larson S.M. Early tumor response to Hsp90 therapy using HER2 PET: comparison with 18F-FDG PET. J Nucl Med. 2006;47(5):793–6. [PMC free article: PMC3193602] [PubMed: 16644749]
38.
Smith-Jones P.M., Solit D.B., Akhurst T., Afroze F., Rosen N., Larson S.M. Imaging the pharmacodynamics of HER2 degradation in response to Hsp90 inhibitors. Nat Biotechnol. 2004;22(6):701–6. [PubMed: 15133471]
39.
Tada H., Higuchi H., Wanatabe T.M., Ohuchi N. In vivo real-time tracking of single quantum dots conjugated with monoclonal anti-HER2 antibody in tumors of mice. Cancer Res. 2007;67(3):1138–44. [PubMed: 17283148]

Views

Search MICAD

Limit my Search:


Related information

  • PMC
    PubMed Central citations
  • PubMed
    Links to PubMed

Similar articles in PubMed

See reviews...See all...

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...