使用斑马鱼检测人血浆、活细胞、人体组织和整个生物体中 DPP IV 的比例双光子近红外探针,ACS Sensors

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使用斑马鱼检测人血浆、活细胞、人体组织和整个生物体中 DPP IV 的比例双光子近红外探针
ACS Sensors ( IF 8.2 ) Pub Date : 2023-02-27 , DOI: 10.1021/acssensors.2c02025
Javier Valverde-Pozo ₁ , Jose M Paredes ₁ , Thomas J Widmann ₂ , Carmen Griñan-Lison _{2,

3,

4} , Giada Ceccarelli ₅ , Antimo Gioiello ₅ , M Eugenia Garcia-Rubiño ₁ , Juan A Marchal _{3,

6,

7} , Jose M Alvarez-Pez ₁ , Eva M Talavera ₁

Affiliation

DPP IV，也称为 CD26 淋巴细胞 T 表面抗原，是一种跨膜糖蛋白，也存在于血液循环中。它在葡萄糖代谢和 T 细胞刺激等多个过程中起着重要作用。此外，它在肾癌、结肠癌、前列腺癌和甲状腺癌组织中过表达。它还可以作为溶酶体贮积病患者的诊断工具。在生理和疾病条件下获得这种酶活性读数的生物学和临床重要性促使我们设计了一种近红外 (NIR) 荧光探针，该探针还具有比率测量和可被两个同时 NIR 光子激发的特性. 该探针包括组装酶识别基团 (Gly-Pro) (Mentlein, 1999; Klemann et al.,H -pyran, DCM-NH ₂ ) 干扰其 NIR 特征内部电荷转移 (ICT) 发射光谱。当 DPP IV 特异性酶促作用释放二肽基团时，供体-受体 DCM-NH ₂得到恢复，形成一个显示高比例荧光输出的系统。有了这个新探针，我们已经能够使用斑马鱼快速有效地检测活细胞、人体组织和整个生物体中 DPP IV 的酶活性。此外，由于有可能被两个光子激发，我们可以避免原始血浆在被可见光激发时产生的自发荧光和随后的光漂白，实现无干扰地检测该介质中DPP IV的活性。

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Ratiometric Two-Photon Near-Infrared Probe to Detect DPP IV in Human Plasma, Living Cells, Human Tissues, and Whole Organisms Using Zebrafish

DPP IV, otherwise known as CD26 lymphocyte T surface antigen, is a transmembrane glycoprotein also found in circulation in the blood. It plays an important role in several processes like glucose metabolism and T-cell stimulation. Moreover, it is overexpressed in renal, colon, prostate, and thyroid human carcinoma tissues. It can also serve as a diagnostic in patients with lysosomal storage diseases. The biological and clinical importance of having readouts for the activity of this enzyme, in physiological and disease conditions, has led us to design a near-infrared (NIR) fluorimetric probe that also has the characteristics of being ratiometric and excitable by two simultaneous NIR photons. The probe consists of assembling an enzyme recognition group (Gly-Pro) (Mentlein, 1999; Klemann et al., 2016) on the two-photon (TP) fluorophore (derivative of dicyanomethylene-4H-pyran, DCM-NH₂) disturbing its NIR characteristic internal charge transfer (ICT) emission spectrum. When the dipeptide group is released by the DPP IV-specific enzymatic action, the donor–acceptor DCM-NH₂ is restored, forming a system that shows high ratiometric fluorescence output. With this new probe, we have been able to detect, quickly and efficiently, the enzymatic activity of DPP IV in living cells, human tissues, and whole organisms, using zebrafish. In addition, due to the possibility of being excited by two photons, we can avoid the autofluorescence and subsequent photobleaching that the raw plasma has when it is excited by visible light, achieving detection of the activity of DPP IV in that medium without interference.

更新日期：2023-02-27

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