光纤生物传感器在HER3抗体药物定量检测中的应用
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摘要
为了实现对生物分子间相互作用过程的实时、灵敏、快速监测,获得生物分子的有无、浓度与相互作用的动力学参数信息,本文设计了基于光纤生物传感器的生物亲和性检测方法。首先,针对光干涉生物亲和性传感检测系统的光学传输系统"Y"型分叉光纤与光纤探针之间的耦合问题,提出了自聚焦透镜与石英光纤耦合结构,该耦合结构偏心公差能够达到0.02 mm,倾斜公差能够达到0.1°;针对干涉光谱信号的高频噪声问题,采用一种改进的经验模态分解干涉光谱信号处理方法,有效避免了干涉光谱曲线滤波处理后极值点位置的偏移;同时采用局部拟合极值点计算生物分子膜层厚度的方法,将生物分子膜层厚度的分辨率提高到50 pm。利用所搭建的光干涉生物亲和性检测系统,建立了HER3-Ig G1抗体药物利用金纳米粒子进行信号放大,实现对其浓度进行定量检测的新方法,检测过程中无需清洗,不产生交叉污染。实验结果表明:系统检测限能达到0.082 6μg/m L,该系统具有检测时间短,测量准确、精度高、成本低廉等特点,能够应用于药代动力学研究中。
In order to realize the real-time,sensitively and rapid monitoring of the interactions between biomolecules,and obtain kinetic parameters of biomolecules existence,concentration and interaction,an affinity detection method based on fiber biosensor is designed in this paper.Firstly,a coupling structure between a self-focusing lens and a quartz fiber is proposed for the coupling problem between the optical transmission sys-tem "Y"-type bifurcated optical fiber and the fiber probe in the optical interference bio-affinity sensing system.The coupling structure has an eccentricity tolerance of up to 0.02 mm,and a tilt tolerance of 0.1°.In order to solve the problem of high frequency noise of the interference spectrum,an improved EMD interference spectral signal processing method is adopted to effectively avoid the deviation of the extreme point position after filtering of interference spectral curves; at the same time,method for calculating the thickness of biomolecule film layer by using locally fitted extreme points is adopted to increase the resolution of the biofilm layer thickness to 50 pm.Using the established light interference bio-affinity detection system,a new method for the detection of concentrations of HER3-Ig G1 antibody using gold nanoparticles for signal amplification was established.There was no need for cleaning and no cross-contamination will occur during the detection process.The experimental results show that the detection limit of the system reaches 0.0826μg/ml.Due to the advantages of short detection time,accurate measurement,high precision,and low cost,the system is suitable for the study of pharmacokinetics.
In order to realize the real-time,sensitively and rapid monitoring of the interactions between biomolecules,and obtain kinetic parameters of biomolecules existence,concentration and interaction,an affinity detection method based on fiber biosensor is designed in this paper.Firstly,a coupling structure between a self-focusing lens and a quartz fiber is proposed for the coupling problem between the optical transmission sys-tem "Y"-type bifurcated optical fiber and the fiber probe in the optical interference bio-affinity sensing system.The coupling structure has an eccentricity tolerance of up to 0.02 mm,and a tilt tolerance of 0.1°.In order to solve the problem of high frequency noise of the interference spectrum,an improved EMD interference spectral signal processing method is adopted to effectively avoid the deviation of the extreme point position after filtering of interference spectral curves; at the same time,method for calculating the thickness of biomolecule film layer by using locally fitted extreme points is adopted to increase the resolution of the biofilm layer thickness to 50 pm.Using the established light interference bio-affinity detection system,a new method for the detection of concentrations of HER3-Ig G1 antibody using gold nanoparticles for signal amplification was established.There was no need for cleaning and no cross-contamination will occur during the detection process.The experimental results show that the detection limit of the system reaches 0.0826μg/ml.Due to the advantages of short detection time,accurate measurement,high precision,and low cost,the system is suitable for the study of pharmacokinetics.
引文
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[15]程知,何枫,张巳龙,等.趋势项调制的小波EMD联合方法用于湍流激光雷达r0廓线去噪[J].光学学报,2017,37(12):1201002.CHENG ZH,HE F,ZHANG S L,et al..Trend modulated combination method of wavelet and EMD for r0 profile of turbulence lidar[J].Acta Optica Sinica,2017,37(12):1201002.(in Chinese)
[16]李欣,梅德庆,陈子辰.基于经验模态分解和希尔伯特-黄变换的精密孔镗削颤振特征提取[J].光学精密工程,2011,19(6):1291-1297.LI X,MEI D Q,CHEN Z CH.Feature extraction of chatter for precision hole boring processing based on EMD and HHT[J].Opt.Precision Eng.,2011,19(6):1291-1297.(in Chinese)
[2]POLLET J,DELPORT F,JANSSEN K,et al..Fiber optic SPR biosensing of DNA hybridization and DNA-protein interactions[J].Biosensors&Bioelectronics,2009,25(4):864-869.
[3]米永巍,刘国华,张维.SPR传感技术在生物医学工程中的研究与应用[J].医疗卫生装备,2008,29(6):31-34.MI Y W,LIU G H,ZHANG W.Research and application of SPR sensor technique in biomedical engineering[J].Chinese Medical Equipment Journal,2008,29(6):31-34.(in Chinese)
[4]POLLET J,DELPORT F,JANSSEN K P F,et al..Fiber optic SPR biosensing of DNA hybridization and DNA-protein interactions[J].Biosensors and Bioelectronics,2009,25(4):864-869.
[5]鲁丁强,庞广昌.电化学纳米免疫传感器在食品安全检测中的应用展望[J].食品科学,2014,35(8):6-10.LU D Q,PANG G CH.Recent development and application of electrochemical nanometer immunosensors in food detection[J].Food Science,2014,35(8):6-10.(in Chinese).
[6]刘春春,杭海英.生物大分子相互作用检测技术新进展——三色荧光级联荧光共振能量转移技术[J].生物化学与生物物理进展,2006(3):292-296.LIU CH CH,HANG H Y.A progress in detection of interactions between macromolecules:linked FRET using three color fluorophore[J].Progress in Biochemistry and Biophysics,2006(3):292-296.(in Chinese).
[7]胡耀明,梁大开,张伟,等.基于强度调制的光纤SPR系统的研究[J].压电与声光,2009,31(1):44-46.HU Y M,LIANG D K,ZHANG W.Research on the light intensity-modulation optic-fiber SPR system[J].Piezoelectectrics&Acoustooptics,2009,31(1):44-46.(in Chinese).
[8]CONCEPCION J,WITTE K,WARTCHOW C,et al..Label-free detection of biomolecular interactions using biolayer interferometry for kinetic characterization[J].Combinatorial Chemistry&High Throughput Screening,2009,12(8):791-800.
[9]唐春晓,李恩邦,王长乐,等.基于多模干涉的反射式光纤生物传感器的研究[J].光电子·激光,2011,22(8):1138-1142.TANG CH X,LI E B,WANG CH L.Reflection-type fiber-optic biosensor based on multimode interference[J].Journal of Optoelectronics·Laser,2009,12(8):791-800.(in Chinese)
[10]沃江海.高精度干涉型光纤传感器的理论与实验研究[D].武汉:华中科技大学,2014.WO J H.Theoretical and experimental investigation on the high sensitive interferometric fiber-optic sensors[D].Wuhan:Graduate University of the Huazhong University of Science&Technology,2014.(in Chinese)
[11]王驰,许婷婷,毕书博,等.测量自聚焦光纤透镜聚焦常数的曲线拟合算法[J].光学精密工程,2015,23(12):3309-3315.WANG CH,XU T T,BI SH B,et al..Curve-fitting algorithm of measuring focusing constant of gradient-index fiber lens[J].Opt.Precision Eng.,2015,23(12):3309-3315.(in Chinese)
[12]王延东,杨春雷,董文辉.SINS初始对准中光纤陀螺EMD滤波[J].中国光学,2015,8(6):934-940.WANG Y D,YANG CH L,DONG W H.EMD filtering of fiber gyro in initial alignment of SINS[J].Chinese Optics,2015,8(6):934-940.(in Chinese)
[13]王婷.EMD算法研究及其在信号去噪中的应用[D].哈尔滨:哈尔滨工程大学,2010.WANG T.Research on EMD algorithm and its application in signal denoising[D].Haerbin:Graduate University of the Haerbin Engineering University,2010(in Chinese)
[14]潘潘,席丽霞,张晓光,等.基于经验模态分解的偏振模色散测量实验研究[J].光学学报,2017,45(1):0106002.PANG P,XI L X,ZHANG X G,et al..Experimental research on polarization mode dispersion measurement based on empirical mode decomposition[J].Acta Optica Sinica,2017,45(1):0106002.(in Chinese)
[15]程知,何枫,张巳龙,等.趋势项调制的小波EMD联合方法用于湍流激光雷达r0廓线去噪[J].光学学报,2017,37(12):1201002.CHENG ZH,HE F,ZHANG S L,et al..Trend modulated combination method of wavelet and EMD for r0 profile of turbulence lidar[J].Acta Optica Sinica,2017,37(12):1201002.(in Chinese)
[16]李欣,梅德庆,陈子辰.基于经验模态分解和希尔伯特-黄变换的精密孔镗削颤振特征提取[J].光学精密工程,2011,19(6):1291-1297.LI X,MEI D Q,CHEN Z CH.Feature extraction of chatter for precision hole boring processing based on EMD and HHT[J].Opt.Precision Eng.,2011,19(6):1291-1297.(in Chinese)