Bi-photon imaging and diagnostics using ultra-small diagnostic probes engineered from semiconductor nanocrystals and single-domain antibodies

Hilal Hafian; Alyona Sukhanova; Patrick Chames; Daniel Baty; Michel Pluot; Jacques H. M. Cohen; Igor R. Nabiev; Jean-Marc Millot

doi:10.1117/12.960493

24 October 2012 Bi-photon imaging and diagnostics using ultra-small diagnostic probes engineered from semiconductor nanocrystals and single-domain antibodies

Hilal Hafian, Alyona Sukhanova, Patrick Chames, Daniel Baty, Michel Pluot, Jacques H. M. Cohen, Igor R. Nabiev, Jean-Marc Millot

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Proceedings Volume 8548, Nanosystems in Engineering and Medicine; 85480R (2012) https://doi.org/10.1117/12.960493
Event: SPIE Nanosystems in Engineering + Medicine, 2012, Incheon, Korea, Republic of

Abstract

Semiconductor fluorescent quantum dots (QDs) have just demonstrated their numerous advantages over organic dyes in bioimaging and diagnostics. One of characteristics of QDs is a very large cross section of their twophoton absorption. A common approach to biodetection by means of QDs is to use monoclonal antibodies (mAbs) for targeting. Recently, we have engineered ultrasmall diagnostic nanoprobes (sdAb–QD) based on highly oriented conjugates of QDs with the single-domain antibodies (sdAbs) against cancer biomarkers. With a molecular weight of only 13 kDa (12-fold smaller than full-size mAbs) and extreme stability and capacity to refolding, sdAbs are the smallest functional Ab fragments capable of binding antigens with affinities comparable to those of conventional Abs. Ultrasmall diagnostic sdAb–QD nanoprobes were engineered through oriented conjugation of QDs with sdAbs. This study is the first to demonstrate the possibility of immunohistochemical imaging of colon carcinoma biomarkers with sdAb–QD conjugates by means of two-photon excitation. The optimal excitation conditions for imaging of the markers in clinical samples with sdAb–QD nanoprobes have been determined. The absence of sample autofluorescence significantly improves the sensitivity of biomarker detection with the use of the two-photon excitation diagnostic setup.

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Hilal Hafian, Alyona Sukhanova, Patrick Chames, Daniel Baty, Michel Pluot, Jacques H. M. Cohen, Igor R. Nabiev, and Jean-Marc Millot "Bi-photon imaging and diagnostics using ultra-small diagnostic probes engineered from semiconductor nanocrystals and single-domain antibodies", Proc. SPIE 8548, Nanosystems in Engineering and Medicine, 85480R (24 October 2012); https://doi.org/10.1117/12.960493

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KEYWORDS

Tissues

Diagnostics

Luminescence

Absorption

Nanoprobes

Semiconductors

Nanocrystals

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