基于量子点荧光猝灭法的蛋白质检测芯片
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摘要
基于羧基量子点的荧光猝灭效应,提出了一种检测神经生长因子(NGF)的新方法。研究结果表明,在pH7.5的缓冲溶液中,NGF抗原与羧基量子点标记的NGF抗体发生特异性结合,能使羧基量子点的荧光发生猝灭。在1～20ng·mL~(-1)范围内,荧光猝灭程度与NGF抗原质量浓度呈良好的线性关系,线性系数为0.9696,检测下限为1ng·mL~(-1)。所提检测方法具有良好的抗干扰特性,在诊断检测方面具有巨大的应用潜力。
A new method for detecting the nerve growth factors(NGF)is proposed based on the fluorescence quenching effect of carboxyl quantum dots.The research results show that in the pH7.5 buffer solution,the NGF antigen specifically binds to the carboxyl quantum dot-labeled NGF antibody,which can quench the fluorescence of the carboxyl quantum dots.The fluorescence quenching degree and the NGF antigen mass concentration has a good linear relationship in the range of 1-20 ng/mL,and the linear correlation coefficient is 0.9696 with the detection limit of 1 ng/mL.This proposed method has good anti-interference characteristics and has great potential applications in diagnosis and detection.
A new method for detecting the nerve growth factors(NGF)is proposed based on the fluorescence quenching effect of carboxyl quantum dots.The research results show that in the pH7.5 buffer solution,the NGF antigen specifically binds to the carboxyl quantum dot-labeled NGF antibody,which can quench the fluorescence of the carboxyl quantum dots.The fluorescence quenching degree and the NGF antigen mass concentration has a good linear relationship in the range of 1-20 ng/mL,and the linear correlation coefficient is 0.9696 with the detection limit of 1 ng/mL.This proposed method has good anti-interference characteristics and has great potential applications in diagnosis and detection.
引文
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[2]Colangelo A M,Bianco M R,Vitagliano L,et al.Anew nerve growth factor-mimetic peptide active on neuropathic pain in rats[J].Journal of Neuroscience,2008,28(11):2698-2709.
[3]Coskun S,Varol S,Ozdemir H H,et al.Association of brain-derived neurotrophic factor and nerve growth factor gene polymorphisms with susceptibility to migraine[J].Neuropsychiatric Disease&Treatment,2016,12(1):1779-1785.
[4]Aloe L,Rocco M L,Balzamino B O,et al.Nerve growth factor:Role in growth,differentiation and controlling cancer cell development[J].Journal of Experimental&Clinical Cancer Research,2016,35:116-122.
[5]Bonini S,Lambiase A,Rama P,et al.Phase I trial of recombinant human nerve growth factor for neurotrophic keratitis[J].Ophthalmology,2018,125(9):1468-1471.
[6]Ketschek A,Jones S,Spillane M,et al.Nerve growth factor promotes reorganization of the axonal microtubule array at sites of axon collateral branching[J].Developmental Neurobiology,2015,75(12):1441-1461.
[7]Medina-Sánchez M,Miserere S,Merko9i A.Nanomaterials and lab-on-a-chip technologies[J].Lab on a Chip,2012,12(11):1932-1943.
[8]Peng M,Bai Z C,Li X J,et al.Controlling widespectrum fluorescence on Au/ZnSe multiheterojunction[J].Applied Physics A,2018,124(7):480.
[9]Bai Z C,Huang Z L,Hao L C,et al.Identifying defects in thin film of high power laser lens by using near field microimaging method[J].Chinese Journal of Lasers,2017,44(1):0103001.白忠臣,黄兆岭,郝礼才,等.近场显微成像法识别高功率激光镜片薄膜内部缺陷[J].中国激光,2017,44(1):0103001.
[10]Chen W B,Ma H,Ye J X,et al.Research progress on quantum dot light emitting diodes[J].Laser&Optoelectronics Progress,2017,54(11):110003.陈雯柏,马航,叶继兴,等.量子点发光二极管的研究进展[J].激光与光电子学进展,2017,54(11):110003.
[11]Jamieson T,Bakhshi R,Petrova D,et al.Biological applications of quantum dots[J].Biomaterials,2007,28(31):4717-4732.
[12]Zhao F,Kim J.Fabrication of a dopamine sensor based on carboxyl quantum dots[J].Journal of Nanoscience and Nanotechnology,2015,15(10):7871-7875.
[13]Lin J Q,Ruan Q Y,Chen G N,et al.Research progress of surface enhanced Raman spectroscopy for cancer detection[J].Laser&Optoelectronics Progress,2013,50(8):080020.林居强,阮秋咏,陈冠楠,等.基于表面增强拉曼散射技术的癌症检测研究进展[J].激光与光电子学进展,2013,50(8):080020.
[14]Yang S M,Yao H,Zhang D P,et al.Droplet-based dielectrophoresis device for on-chip nanomedicine fabrication and improved gene delivery efficiency[J].Microfluidics and Nanofluidics,2015,19(1):235-243.
[15]Mark D,Haeberle S,Roth G,et al.Microfluidic lab-on-a-chip platforms:Requirements,characteristics and applications[J].Chemical Society Reviews,2010,39(3):1153-1182.
[16]Sangamithirai D,Munusamy S,Narayanan V,et al.Fabrication of neurotransmitter dopamine electrochemical sensor based on poly(o-anisidine)/CNTs nanocomposite[J].Surfaces and Interfaces,2016,4:27-34.
[17]Gu B,Sheng X,Ye Z C.Research on quantum dot apply to LCD backlight[J].Laser&Optoelectronics Progress,2015,52(2):022201.顾宝,盛欣,叶志成.量子点应用于液晶显示背光的研究[J].激光与光电子学进展,2015,52(2):022201.
[18]Jokerst J V,Jacobson J W,Bhagwandin B D,et al.Programmable nano-bio-chip sensors:Analytical meets clinical[J].Analytical Chemistry,2010,82(5):1571-1579.
[19]Baird C L,Myszka D G.Current and emerging commercial optical biosensors[J].Journal of Molecular Recognition,2001,14(5):261-268.
[20]Dunbar S A,Vander Zee C A,Oliver K G,et al.Quantitative,multiplexed detection of bacterial pathogens:DNA and protein applications of the Luminex LabMAPTM system[J].Journal of Microbiological Methods,2003,53(2):245-252.
[21]Croft H,Malinowski T,Krizbai L,et al.Use of Luminex xMAP-derived Bio-Plex bead-based suspension array for specific detection of PPV W and characterization of epitopes on the coat protein of the virus[J].Journal of Virological Methods,2008,153(2):203-213.
[22]Chen F T.Research of diagnosis for early beast carcinoma by surface enhanced laser desorption/ionization-time of flight-mass spectrometry[D].Shanghai:Second Military Medical University,2009:12-28.陈锋涛.SELDI质谱技术在乳腺癌早期诊断中的应用研究[D].上海:第二军医大学,2009:12-28.
[23]Cui D F,Wang Y J,Cai H Y,et al.Detection of 6-oxyguanine-DNA methyltransferase by SPRBiochemical Analyzer.[J].Progress in Natural Science,2003,13(8):874-876.崔大付,王于杰,蔡浩原,等.SPR生化分析仪对6-氧甲基鸟嘌呤-DNA甲基转移酶的检测[J].自然科学进展,2003,13(8):874-876.
[24]Niu Y.Research on induction biology system and application of elliptical imaging protein chip biosensor[D].Beijing:University of Chinese Academy of Sciences,2011:22-48.牛宇.椭偏成像蛋白质芯片生物传感器感应生物学系统和应用研究[D].北京:中国科学院大学,2011:22-48.
[25]Zhang Y,Bai Z C,Huang Z L,et al.Influence of distance between CdSe quantum dot and gold nanoparticle on system fluorescence[J].Laser&Optoelectronics Progress,2018,55(7):072601.张莹,白忠臣,黄兆岭,等.金纳米粒子与CdSe量子点间的距离对体系荧光的影响[J].激光与光电子学进展,2018,55(7):072601.
[26]Xiao F Z.Preparation and application of biochip carriers based on nano-membranes[D].Hengyang:University of South China,2008:5-20.肖方竹.纳米膜基底型生物芯片载体制备及应用研究[D].衡阳:南华大学,2008:5-20.