雌二醇的水凝胶毛细管电泳免疫分析

雌二醇是雌性激素中最重要和作用最强的一种激素 .临床上 ,雌二醇的长期监测可用于研究与激素相关的致癌物作用机理、绝经期妇女的雌激素补充及不孕病人的跟踪治疗等方面 .研究发现 ,人体中雌二醇含量与某些肿瘤如乳腺癌、子宫癌和肝癌等密切相关 [1] .由于雌二醇在体液中含量很低 ,难于测定 .免疫分析法的出现给雌二醇的测定带来了变革 ,提高了测定的灵敏度和特异性 .目前测定雌二醇的免疫分析方法有放射免疫分析法 ( RIA) [2 ]、酶免疫分析法 ( EIA) [3～ 7]、荧光免疫分析法 ( FIA) [8]和化学发光与生物发光免疫分析法 [9,10 ] .但 R…     

热敏相分离荧光免疫分析乙肝表面抗原的新方法

以N-异丙基丙烯酰胺为单体,将水溶性热敏高分子聚异丙基丙烯酰胺(PNIP)和抗体偶联,用异硫氰酸荧光素标记羊抗人乙肝表面抗原抗体,建立了夹心型热敏相分离荧光免疫分析乙肝表面抗原(HBsAg)的新方法.评价了抗体在PNIP上的固定效率和非特异性吸附情况.HBsAg在0.5～100μg/mL范围内与体系荧光强度呈良好的线性关系,检出限为10ng/mLHBsAg.该方法既具有均相免疫分析的快速性,又有固相免疫分析的高灵敏度.用于乙肝病人血清中HBsAg水平的测定,结果令人满意.     

具有温度敏感和智能荧光特性的聚合物包覆金纳米粒子的研究

以4-二甲氨基吡啶稳定的金纳米粒子(DMAP-AuNPs)为前驱体,通过与端基含二甲氨基查尔酮的聚N-异丙基丙烯酰胺(PNIPAM-DMAC)进行配体置换,制备了具有温度敏感和智能荧光特性的PNIPAM-DMAC聚合物包覆的金纳米粒子(PNIPAM-AuNPs),并通过紫外可见吸收光谱、透射微镜、核磁共振氢谱、红外光谱和热重分析仪对其形貌和结构进行了分析和表征.PNIPAM-AuNPs能够很好地分散在水、乙醇、丙酮、氯仿和四氢呋喃中,其表面等离子共振(SPR)吸收特征波长分别为537、527.6、527.4、532.2和530.4 nm.PNIPAM-AuNPs的表面聚合物接枝密度为1～2条聚合物链/nm2.温敏性结果表明,PNIPAM-AuNPs具有低临界溶解温度(LCST),其LCST温度比相应的PNIPAM-DMAC聚合物低2.5 K;在PNIPAM-AuNPs发生相转变前后,金纳米粒子的SPR特征峰由537 nm蓝移至525 nm处.荧光性能研究表明,随着溶剂极性的增加,PNIPAM-AuNPs 的荧光特征波长发生红移,荧光强度先增加后减小,具有极性敏感的荧光特性;在25℃时,PNIPAM-AuNPs水溶液几乎无荧光,而当温度升高到45℃后,荧光强度显著提高,表现出温度“开/关”的荧光特性.     

时间分辨荧光免疫分析法间接测定雌二醇

以氯磺酰基噻吩甲酰三氟丙酮（CTTA）为铕（Eu)的螯合剂,羊抗鼠（SAM）的IgG为二抗,用SAM－IgG-CTTA-Eu作标记二抗,建立了以竞争抑制为基础的时间分辨荧光免疫分析测定离雌二醇（E2）的新方法。同均相方法相比灵敏度有很大提高,测定雌二醇（E2）的线性范围为2．5－200pg/mL,检测限为2.5pg/mL。这一方法可望用于E2的临床检测。     

时间分辨荧光免疫分析法直接测定雌二醇

合成了雌二醇抗原,并获得了雌二醇单克隆抗体,通过酶联免疫吸附法测得效价为１．０×１０＾７。使用洗脱增强－时间分辨荧光免疫分析法测定了血清中的雌二醇,浓度范围为１．０×１０＾－２－１．０×１０＾３μｇ／Ｌ,最低检出限为５．６ｎｇ／Ｌ,批内ＲＳＤ％小于３．７％,批间ＲＳＤ％小于７．５％。将测定结果 放射免疫分析法作了比较。     

N-异丙基丙烯酰胺温度敏感性水凝胶相转变动力学研究及其应用

 研究指出表观二级动力学方程可以很好地描述N-异丙基丙烯酰胺水凝胶的溶胀和消溶胀动力学.即溶胀动力学方程为dR/dt=k₁(R_e-R)²,消溶胀动力学方程为-dR/dt=k_c(R-R_e)².把这种水凝胶用于分离高分子水溶液时可引入“单位溶张比分离循环的合理时耗”这样一个参量.它根据溶胀和消溶胀过程中的起始溶胀比、平衡溶胀比、表观溶胀动力学常数和表观消溶胀动力学常数求出.具体公式为△t₁(T_s,T_c)=2/[R_c(T_s)-R₀(T_s)]²k_s(T_s)+15/[R₀(T_c)- R_s(T_c)]²k_c(T_c)^1/2在理想情况下,分离过程的“总合理时耗”与△t_1成正比,比例系数为分离过程中的除水总量与干凝胶用量的比值,即△t_r=W_W/WG·△t₁.当根据二个动力学方程求得的总时耗计算值处于(0.9△t_r,1.1△t_r)范围内时,表明所选干凝胶用量和循环溶胀比区段均合适.     

明胶-聚异丙基丙烯酰胺水凝胶的pH、温度敏感性

用明胶(Gel)和N 异丙基丙烯酰胺(NIPAM)为原料,制备了Gel/聚异丙基丙烯酰胺(PNIPAM)水凝胶;研究了不同含量的水凝胶的温度、pH敏感性。结果表明:温度对水凝胶pH敏感性的影响取决于水凝胶的组成。明胶含量高的水凝胶,其pH敏感性几乎不受温度的影响;当0.500.90时,pH值几乎不影响水凝胶的温敏性。     

温度敏感水凝胶包覆长周期光纤光栅传感器研究

以N-异丙基丙烯酰胺(NIPAM)为单体、N,N'-亚甲基双丙烯酰胺(MBAA)为交联剂、安息香二甲醚(DMPA)为引发剂,利用紫外光引发聚合制备了一系列温度敏感性聚(N-异丙基丙烯酰胺)(PNIPAM)水凝胶,并对其性能进行了测定.结果表明,PNIPAM水凝胶的平衡膨胀比随着交联程度的变化而改变.当交联程度适当时,水凝胶可具有最大的溶胀比.在此研究基础上,利用浸渍提拉法在长周期光纤光栅(LPFG)包层外制备了PNIPAM水凝胶薄膜包覆层.研究了得到的LPFG传感器对温度和湿度的响应性,该类型传感器表现出对温度的灵敏响应性.     

N－异丙基丙烯酰胺系温度敏感聚合物和水凝胶的研究进展

综述了近几年来Ｎ－异丙基丙烯酰胺系温度敏感聚合物和水凝胶研究的最新进展，对其合成方法，性质与结构的关系和开发应用的研究都作了较详尽的报道。     

可控相转变温度热敏高分子的制备及其在免疫分析中的应用

合成了一种新型的快速响应热敏高分子聚N－异丙基丙烯酰胺－丙烯酰胺[P(NIP-co-AA)]，通过改变丙烯酰胺的含量可以改变高分子的临界溶解温度（LCST），使之用于不同用途。其中，将相转变温度（Ttr)在37℃的热敏高分子用于免疫分析的载体，建立了夹心型荧光免疫分析兔IgG的新方法。与聚N－异丙基丙烯酰胺（PNIP）作载体相比，两灵敏度相当，但由于相转变温度的提高，使得免疫反应的温度更接近于生物体的生理环境，并使免疫反应速率得到提高。该方法线性范围为0-1000μg/L；检出限为10μg/L。用于兔血清中兔IgG的测量，结果令人满意。     

Poly-N-Isopropylacrylamide Hydrogel Based Fluoroimmunoassay of Methyltestosterone

IntroductionMethyltestosterone(MT)isasynthesizedsteroid.Athletesusesyntheticsteroidstoincreasetheirsportgrade.This,fromtheviewofethics,violatedtheoriginalrulesofsport.Therefore,themonitoringofthedopeabusehasbecomemoreandmoreimportantworldwide.Sofar.themonitoringiscommonlycarriedoutbychromatographicseparationfollowedbymassspectrometricanalysis.Thetargetsamplesarenormallyurinel~2.Themonitoringbasedonbloodsampleismainlyusedasacomplementarymethodtoovercometheshortcomingfromtheanalysisusingurines…     

Thermally Reversible Hydrogel Based Fluoroimmunoassay of Methyltestosterone

ABSTRACT

A novel immunoassay of methyltestosterone (MT) in serum is developed. We synthesized the antigen of MT (Ag_MT) by covalently bonding it to BSA and raised its antibody (Ab_MT) from rabbit. Ab_MT was conjugated to poly-N-isopropylacrylamide (p-NIPAAm) to form Ab_MT bound thermally reversible hydrogel (p-NIPAAm-Ab_MT). A competitive immunoassay method based on the competition of fluorescein isothiocyanate (FITC) labeled MT antigen (FITC-Ag_MT) and MT with limited amounts of p-NIPAAm-Ab_MT was established. The separation in the assay process was achieved by precipitation of the immuno-complex above its critical solution temperature. The detection limit of MT is 50 ng/ml. The recoveries of MT in human serum are above 93%.     

Development of Polarization Fluoroimmunoassay for the Detection of s-Triazine Herbicides

Abstract

A rapid, non-isotopic polarization fluoroimmunoassay (PFIA) for the monitoring of the simazine (striazine herbicide) level in water was developed. Polyclonal antiserum was raised in rabbits by immunization with simazine – Keyhole Limpet Haemocyanin conjugate. Sensitivity of the PFIA with the use of heterologous tracer with the shortest bridge between antigen and fluorescein proved to be the highest. All analytical criteria for PFIA were satisfied. The detection limit of simazine (3 ng/ml in 50 μl of sample) was comparable to that for liquid or gas chromatography method. The detection limit of ELISA using the same antiserum and conjugate derivative of atrazine with horseradish peroxidase was 0.1 ng/mL of simazine. The cross-reactivity for PFIA with widely used s-triazine herbicides: atrazine, propazine, terbuthylazine was 100%, 32% and 20%, respectively. The cross-reactivity for PFIA with some metabolites of s-triazines and other herbicides was negligible.     

基于适配体和抗体的生物传感器在雌二醇检测中的应用

雌二醇(Estradiol,E2)是一种主要的环境雌激素,具有作用强、危害广等特征,可扰乱人体及动物体的正常内分泌功能,从而产生不利影响,因此E2的快速、准确检测对于食品安全和公共卫生至关重要.传统的检测方法主要是基于大型仪器分析,这些方法具有高灵敏度、准确性的同时也存在操作繁琐、成本较高、仅限于在实验室内进行等缺陷....     

Production of Antibodies and Development of Specific Polarization Fluoroimmunoassay for Acetochlor

Specific polyclonal antibodies towards acetochlor (2-chloro- N -(ethoxymethyl)- N -(2-ethyl-6-methylphenyl)acetamide) were obtained from rabbits immunized against a 3-mercaptopropionic acid derivative of acetochlor, covalently attached to bovine serum albumin. A polarization fluoroimmuoassay (PFIA) based on these antibodies was developed and optimized to detect acetochlor in water samples. The optimized PFIA had a detection limit of 9 µg/L, linear working range from 50 to 5500 µg/L and within-assay coefficient of variation less than 4%. Cross-reactivity studies demonstrated that these antibodies are capable of specific detection of acetochlor amongst structurally related chloroacetanilide herbicides. Assay cross-reactivity values were: alachlor 0%, metolachlor 2.4%, propachlor 0%, butachlor 0.2% and dimethachlor 0.5%. Five organic solvents commonly used in sample extraction were evaluated for their effect on acetochlor PFIA performance, and methanol and ethanol were found to be compatible with the assay up to 10% v/v.     

Hapten Design and Development of Polarization Fluoroimmunoassay for Nonylphenol

Polyclonal antibodies to nonylphenol (NP), raised against two different haptenic derivatives were characterized by a rapid, homogeneous polarization fluoroimmunoassay (PFIA). The first hapten, ω-(4-hydroxyphenyl)nonanoic acid (NP9), was designed to mimic the linear NP isomer and contains hydroxyphenyl and linear alkyl chain moieties of the NP molecule. The second hapten, 4-aminophenol (4AP), contains the hydroxyphenyl moiety of NP molecule alone and thus potentially mimics various phenolic compounds with different side chain structures. A number of fluorescent labeled antigens (tracers) with various structures of the spacer arm between the antigen and the fluorescent dye was synthesized and used to optimize the competitor structure for NP-specific PFIA. The most sensitive assay with limit of detection (LOD) and IC₅₀ values of 8 and 53 mg L^–1, respectively, was obtained when anti-NP9 antibody and NP9-labeled antigen were used. Anti-NP9 resulted in more specific assay, where the cross-reactivity toward the relative phenolic compounds did not exceed 5%. Anti-4AP displayed substantial recognition of several bis-substituted phenols, including 2-amino-4-chlorophenol and 2,4-dinitrophenol.     

Fiber-Optic Sensor with a Sandwich Binding Technique for Fluoroimmunoassay

Abstract

We report the development of a fiber-optic sensor based on the principle of the sandwich binding technique for the fluoroimmunoassay. Anti-mouse immunoglobulin G (IgG) antibody was immobilized on the membrane mounted to the edge of the fiber. The sensor was exposed to the solution containing mouse IgG and then allophycocyanin conjugated anti-mouse IgG antibody was added. The helium-neon (He-Ne) laser provides excitation of the sandwich binding antigen-antibody complex. This results in fluorescence emission at the membrane. Increase in the fluorescence intensity from sensing tip was proportional to the amount of mouse IgG in the sample. A linear relationship was obtained between the fluorescence intensity increase and the mouse IgG concentration in the range 0.3 - 9.0 μg/ml.     

镧系螯合物荧光免疫分析法测定对硫磷

本文应用一种新的镧系螯合物荧光标记法研究在药对硫磷的免疫分析测定，并作了血清、尿和水样品的回收分析，检测下限为１００ｐｇ／ｍＬ，回收率８７—１０２％。     

带活性末端的热敏高分子的制备及其在荧光免疫分析中的应用

合成了带有活性末端的寡聚N-异丙基丙烯酰胺(ONIPAAm),并考察了其温度敏感性质.以ONI-PAAm作为免疫分析的载体与鼠IgG偶联,以四磺基铁酞菁为标记物,以羊抗鼠IgG抗体为分析模型,建立了竞争型热敏相分离荧光免疫分析新系统.羊抗鼠IgG抗体在0～1500ng/mL范围内与体系相对荧光强度呈良好的线性关系,检测限为2ng/mL.