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Actively targeted in vivo multiplex detection of intrinsic cancer biomarkers using biocompatible SERS nanotags.

Dinish US, Balasundaram G, Chang YT, Olivo M - Sci Rep (2014)

Bottom Line: However, nanotags without antibodies showed no detectable signal after 6 hours.This difference could be due to the specific binding of the bioconjugated nanotags to the receptors on the cell surface.Thus, this study establishes SERS nanotags as an ultrasensitive nanoprobe for the multiplex detection of biomarkers and opens up its potential application in monitoring tumor progression and therapy and development into a theranostic probe.

View Article: PubMed Central - PubMed

Affiliation: 1] Singapore Bioimaging Consortium, Agency for Science Technology and Research (A*STAR), 11 Biopolis Way, Singapore 138667 [2].

ABSTRACT
Surface-enhanced Raman scattering (SERS) technique is becoming highly popular for multiplex biosensing due to the 'fingerprint' Raman spectra from every molecule. As a proof-of-concept, we demonstrated the actively targeted multiplex in vitro and in vivo detection of three intrinsic cancer biomarkers - EGFR, CD44 and TGFβRII in a breast cancer model using three multiplexing capable, biocompatible SERS nanoparticles/nanotags. Intra-tumorally injected antibody conjugated nanotags specifically targeting the three biomarkers exhibited maximum signal at 6 hours and no detectable signal at 72 hours. However, nanotags without antibodies showed no detectable signal after 6 hours. This difference could be due to the specific binding of the bioconjugated nanotags to the receptors on the cell surface. Thus, this study establishes SERS nanotags as an ultrasensitive nanoprobe for the multiplex detection of biomarkers and opens up its potential application in monitoring tumor progression and therapy and development into a theranostic probe.

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In vivo multiplex detection in xenograft tumor: (A):- Image of a tumor bearing mouse from test group used in experiment.(B):-SERS spectra from tumor site in a representative control mice showing the peaks at 1120,1175 and 1650 cm−1 from non bioconjugated SERS nanotags corresponding to Cy5, MGITC and Rh6G respectively. Signal intensity is not detectable 6 hours after injection and nanotags gets cleared fast from the body due to the lack of specific binding. (C):- SERS spectra from tumor site in a representative test mice showing the peaks at 1120, 1175 and 1650 cm−1 from the bioconjugated Cy5, MGITC and Rh6G nanotags bound to TGFβRII, CD44 and EGFR biomarker respectively. Due to active targeting, multiplex SERS spectra is observed up to 48 hours followed by the clearance of nanotags from the mouse body by ~72 hours.
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f4: In vivo multiplex detection in xenograft tumor: (A):- Image of a tumor bearing mouse from test group used in experiment.(B):-SERS spectra from tumor site in a representative control mice showing the peaks at 1120,1175 and 1650 cm−1 from non bioconjugated SERS nanotags corresponding to Cy5, MGITC and Rh6G respectively. Signal intensity is not detectable 6 hours after injection and nanotags gets cleared fast from the body due to the lack of specific binding. (C):- SERS spectra from tumor site in a representative test mice showing the peaks at 1120, 1175 and 1650 cm−1 from the bioconjugated Cy5, MGITC and Rh6G nanotags bound to TGFβRII, CD44 and EGFR biomarker respectively. Due to active targeting, multiplex SERS spectra is observed up to 48 hours followed by the clearance of nanotags from the mouse body by ~72 hours.

Mentions: We carried out the multiplex detection of intrinsic biomarkers in vivo by injecting 200 μl of the three bioconjugated SERS nanotags (MGITC, Cy5 and Rh6G in the ratio 1:1:2) into the centre of the tumor on a subcutaneous MDA-MB-231 breast cancer xenograft mouse model (Fig. 4A). SERS nanotags conjugated to antibodies served as test formulation while non-conjugated nanotags served as control formulation. SERS measurements were taken at regular intervals starting immediately after the injection. The spectra clearly revealed the distinct Raman peaks corresponding to each of the three nanotags from both the control and test mice with least spectral overlap in the 1000–1800 cm−1 region (Fig. 4B and 4C). Raman shift at 1120 cm−1, 1175 cm−1 and 1650 cm−1 corresponds to the Cy5, MGITC and Rh6G respectively.


Actively targeted in vivo multiplex detection of intrinsic cancer biomarkers using biocompatible SERS nanotags.

Dinish US, Balasundaram G, Chang YT, Olivo M - Sci Rep (2014)

In vivo multiplex detection in xenograft tumor: (A):- Image of a tumor bearing mouse from test group used in experiment.(B):-SERS spectra from tumor site in a representative control mice showing the peaks at 1120,1175 and 1650 cm−1 from non bioconjugated SERS nanotags corresponding to Cy5, MGITC and Rh6G respectively. Signal intensity is not detectable 6 hours after injection and nanotags gets cleared fast from the body due to the lack of specific binding. (C):- SERS spectra from tumor site in a representative test mice showing the peaks at 1120, 1175 and 1650 cm−1 from the bioconjugated Cy5, MGITC and Rh6G nanotags bound to TGFβRII, CD44 and EGFR biomarker respectively. Due to active targeting, multiplex SERS spectra is observed up to 48 hours followed by the clearance of nanotags from the mouse body by ~72 hours.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3921631&req=5

f4: In vivo multiplex detection in xenograft tumor: (A):- Image of a tumor bearing mouse from test group used in experiment.(B):-SERS spectra from tumor site in a representative control mice showing the peaks at 1120,1175 and 1650 cm−1 from non bioconjugated SERS nanotags corresponding to Cy5, MGITC and Rh6G respectively. Signal intensity is not detectable 6 hours after injection and nanotags gets cleared fast from the body due to the lack of specific binding. (C):- SERS spectra from tumor site in a representative test mice showing the peaks at 1120, 1175 and 1650 cm−1 from the bioconjugated Cy5, MGITC and Rh6G nanotags bound to TGFβRII, CD44 and EGFR biomarker respectively. Due to active targeting, multiplex SERS spectra is observed up to 48 hours followed by the clearance of nanotags from the mouse body by ~72 hours.
Mentions: We carried out the multiplex detection of intrinsic biomarkers in vivo by injecting 200 μl of the three bioconjugated SERS nanotags (MGITC, Cy5 and Rh6G in the ratio 1:1:2) into the centre of the tumor on a subcutaneous MDA-MB-231 breast cancer xenograft mouse model (Fig. 4A). SERS nanotags conjugated to antibodies served as test formulation while non-conjugated nanotags served as control formulation. SERS measurements were taken at regular intervals starting immediately after the injection. The spectra clearly revealed the distinct Raman peaks corresponding to each of the three nanotags from both the control and test mice with least spectral overlap in the 1000–1800 cm−1 region (Fig. 4B and 4C). Raman shift at 1120 cm−1, 1175 cm−1 and 1650 cm−1 corresponds to the Cy5, MGITC and Rh6G respectively.

Bottom Line: However, nanotags without antibodies showed no detectable signal after 6 hours.This difference could be due to the specific binding of the bioconjugated nanotags to the receptors on the cell surface.Thus, this study establishes SERS nanotags as an ultrasensitive nanoprobe for the multiplex detection of biomarkers and opens up its potential application in monitoring tumor progression and therapy and development into a theranostic probe.

View Article: PubMed Central - PubMed

Affiliation: 1] Singapore Bioimaging Consortium, Agency for Science Technology and Research (A*STAR), 11 Biopolis Way, Singapore 138667 [2].

ABSTRACT
Surface-enhanced Raman scattering (SERS) technique is becoming highly popular for multiplex biosensing due to the 'fingerprint' Raman spectra from every molecule. As a proof-of-concept, we demonstrated the actively targeted multiplex in vitro and in vivo detection of three intrinsic cancer biomarkers - EGFR, CD44 and TGFβRII in a breast cancer model using three multiplexing capable, biocompatible SERS nanoparticles/nanotags. Intra-tumorally injected antibody conjugated nanotags specifically targeting the three biomarkers exhibited maximum signal at 6 hours and no detectable signal at 72 hours. However, nanotags without antibodies showed no detectable signal after 6 hours. This difference could be due to the specific binding of the bioconjugated nanotags to the receptors on the cell surface. Thus, this study establishes SERS nanotags as an ultrasensitive nanoprobe for the multiplex detection of biomarkers and opens up its potential application in monitoring tumor progression and therapy and development into a theranostic probe.

Show MeSH
Related in: MedlinePlus