Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan, 430072, China.
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan, 430072, China.
Anal Chim Acta. 2021 May 29;1161:338479. doi: 10.1016/j.aca.2021.338479. Epub 2021 Apr 5.
Matrix metalloproteinase 2 (MMP2) plays an important role in tumor growth, invasion and metastasis. In this work, a dual-functional magnetic microsphere probe was designed for ICP-MS quantification and fluorescence imaging of MMP2 in cell secretion. In the designed probe, a NH-peptide (-FAM)-biotin was used as a bridge for the combination of carboxylated magnetic beads (MBs-COOH) and streptavidin functionalized gold nanoparticle (Au NP-SA). Initially, the fluorescence of FAM was quenched by Au NP. Since the NH-peptide (-FAM)-biotin had a MMP2-specifically recognized sequence, the peptide was specifically cleaved in the presence of MMP2, thus releasing Au NP for the ICP-MS quantification of MMP2 and turning on the fluorescence of FAM for the fluorescence imaging of MMP2. Under the optimal experimental conditions, a linear range of 0.05-50 ng mL and a limit of detection of 0.02 ng mL were obtained for MMP2. The relative standard deviation (n = 7, c = 0.1 ng mL) of the proposed method was 5.4%. With good sensitivity and good accuracy, the proposed method realized the quantification and imaging of MMP2 in A549 cell secretion. The proposed method was applied to monitor the expression of MMP2 in the A549 cell secretion under the stimulation of Cd, providing a new detection strategy in the study of MMP2-related life process.
基质金属蛋白酶 2(MMP2)在肿瘤生长、侵袭和转移中发挥重要作用。在这项工作中,设计了一种双重功能的磁性微球探针,用于 ICP-MS 定量和细胞分泌中 MMP2 的荧光成像。在设计的探针中,NH-肽(-FAM)-生物素被用作连接羧基化磁性珠(MBs-COOH)和链霉亲和素功能化金纳米粒子(Au NP-SA)的桥梁。最初,FAM 的荧光被 Au NP 猝灭。由于 NH-肽(-FAM)-生物素具有 MMP2 特异性识别序列,因此在存在 MMP2 的情况下,肽被特异性切割,从而释放 Au NP 用于 MMP2 的 ICP-MS 定量,并开启 FAM 的荧光用于 MMP2 的荧光成像。在最佳实验条件下,MMP2 的线性范围为 0.05-50ngmL,检出限为 0.02ngmL。该方法的相对标准偏差(n=7,c=0.1ngmL)为 5.4%。该方法具有良好的灵敏度和准确性,实现了 A549 细胞分泌中 MMP2 的定量和成像。该方法被应用于监测 Cd 刺激下 A549 细胞分泌中 MMP2 的表达,为 MMP2 相关生命过程的研究提供了一种新的检测策略。