温敏抗蛋白吸附共聚物涂层及近红外二区荧光成像分子的设计研究

【作者】 王芳；

【导师】 申有青；

【作者基本信息】 青岛大学 ， 材料工程（专业学位）， 2021， 硕士

【摘要】 蛋白质的非特异性吸附问题涉及到多个研究领域,近年,分析仪器在蛋白质组学、代谢组学、生物标记物等方面取得重大进展,在进行生化分析时,分析物与色谱柱表面的相互作用会对分离效率和迁移时间造成影响,降低系统的分离效果。而涂层可有效解决分析物在色谱柱内壁的吸附问题,提高系统的分离性能,更具分析优势。基于以上背景,本研究以聚乙二醇(PEO)、N-异丙基丙烯酰胺(NIPAAm)、甲基丙烯酸磺酸丙酯(SPMAP)为原料,通过原子转移自由基共聚(ATRP)获得具有双重特性的三嵌段共聚物PEO-PNIPAAm-PSPMAP。该共聚物通过光敏剂重氮树脂(DR)共价键合于疏水性基底并应用于分析化学。共聚物中的亲水嵌段提供了抗蛋白质吸附的基础,NIPAAm嵌段为材料的自清洁提供了可能性,并通过荧光蛋白的动态吸附试验和电泳分析证实了涂层的实用性。通过赋予涂层环境响应自清洁性或再生能力,在改进毛细管涂层的制备方法、提高所制备毛细管涂层的寿命、质量和性能、丰富毛细管涂层的种类等方面具有重要意义,也会对功能化毛细管涂层在蛋白质、多肽等生物分子的分离和微量制备以及质谱联用分析等方面的应用产生积极深远的影响。近红外荧光成像可克服可见光在穿透深度和对比度上的缺陷,大大降低组织的自发荧光和散射,为无创体内生物成像提供了一个更具优势的平台,因此近红外荧光材料,特别是近红外II荧光成像材料成为探查生物组织或器官的研究热点。而相较于其他无机及聚合物的荧光分子来说,荧光小分子的发光性能、生物相容性和生物安全性等性能可通过合理化的结构设计来控制改进,因此具有更大的研究应用潜力。本研究以苯并二噻二唑(BBTD)、3,4-乙烯二氧噻吩(EDOT)、环戊联噻吩(CPDT)为原料通过偶联反应得到供体单元-受体单元-供体单元(D-A-D)的双供体的荧光分子。其中,BBTD可以改善电子的离域作用,CPDT增加分子的共平面性和共轭结构,增加分子的摩尔消光系数,提升吸光效率。EDOT可修饰整个结构,增加空穴的传输,降低分子带隙。该荧光分子的吸收峰和发射峰分都在近红外二区范围,别为900 nm和1120 nm,具有潜在更好的成像特点,利用PSMPEG两亲嵌段形成的纳米颗粒的量子产率达到了0.98%,穿透深度可达到15mm,且该分子可实现肾脏和肝脏代谢,但限于其纳米颗粒尺寸影响,肝脏代谢占主要。后期可经修饰更好的实现其体内代谢和肿瘤靶向富集,提高其在医学应用中的潜力。此类D-A-D型有机小分子经过亲水性修饰提高其生物相容性、靶向或标志物修饰提高其体内特异性定位,抗癌药物或活性物质修饰提高其治疗效果等方式进一步提高其在生物体内的应用潜力,实现诊疗一体化,更好的指导手术,提高其临床应用。更多还原

【Abstract】 The non-specific adsorption of proteins involves many research fields.In recent years,capillary electrophoresis has made great progress in proteomics,metabonomics,biomarkers and so on.In biochemical analysis,the interaction between analyte and column surface will affect the separation efficiency and migration time,and reduce the separation effect of the system.The coating can effectively solve the adsorption problem of analytes on the inner wall of the chromatographic column,improve the separation performance of the system,and has more analytical advantages.Based on the above background,we obtained a triblock copolymer peo-pnipaampspmap by atom transfer radical polymerization(ATRP).The copolymer was covalently bonded to hydrophobic substrate by photosensitizer diazoresin(DR)and applied to analytical chemistry.Sufficient hydrated groups(ether bond,amide group and sulfonic group)in the copolymers provide the basis for the adsorption performance of proteins.The conformation changes caused by the interaction of hydrophilic group and hydrophobic group(isopropyl)at different temperatures provide the possibility for the elastic self-cleaning property of the material,and can prolong the cycle life of the coating material.The antifouling performance and elastic self-cleaning function of the coating were evaluated by dynamic adsorption experiment of fluorescent protein.The coating is used for capillary electrophoresis(CE)coated column,and its excellent protein separation spectrum proves the practicability of the coating.Near infrared fluorescence imaging can greatly reduce the spontaneous fluorescence and scattering of tissues,overcome the defects of visible light in penetration depth and contrast,and provide a more advantageous platform for noninvasive biological imaging in vivo.Therefore,near infrared fluorescent materials,especially near infrared II fluorescent imaging materials,have become a research hotspot in the exploration of biological tissues/organs.Compared with other inorganic and polymer fluorescent molecules,the luminescent properties,biocompatibility and biosafety of small fluorescent molecules can be controlled and improved by reasonable structural design,so it has greater research and application potential.We use donor acceptor donor unit(D-A-D)structure design.For acceptor group,benzothiadiazole(BBTD)has a significant quinone type characteristic,which can improve the electron delocalization,and is the most popular choice.The donor,Cyclopentadithiophene(CPDT)and 3,4-ethylene dioxythiophene(EDOT)can improve the molar extinction coefficient,increase the conjugation performance,and facilitate hole transmission,reduce the band gap and improve the fluorescence performance.The introduction of alkyl chains into the molecule increases solubility.The absorption and emission peaks of the fluorescent molecule are 900 nm and 1120 nm,respectively.The quantum yield of the water-soluble nanoparticles formed by PS-MPEG is 0.98%,and its penetration depth can reach 15 mm,which has good biocompatibility.This kind of small organic molecules can be modified to increase their clinical translation,and then cooperate with biological imaging system to realize living biological imaging therapy and better guide surgery.更多还原