免疫分析法进展

本文首先从试剂分析的观点对免及分析的原理进行了论.述,对抗体作为分析试剂进行了评价并且总结了标记免投分析的三种方式和四个环节;其次,对免疫分析的现状作了综述,并评述了当代免反分析的五个热点:基因工程抗体,生物素一亲合素多重标记体系,时间分辨荧光免疫分析,多组分免疫分析和自动化免疫分析;最后,对免疫分析的发展趋势作了讨论。     

X-射线荧光光谱分析技术的发展

归纳了X-射线荧光光谱分析技术发展的进程。从现代控制技术的改善、仪器检测性能的提高、元素检测范围的扩大等8方面阐述了波长色散X-射线荧光光谱技术的进展,还就能量色散X-射线荧光光谱仪的X射线管和探测器技术的快速发展及近10年来我国在X-射线荧光光谱分析方法方面的论文发表情况进行了总结,对近年来X-射线荧光光谱仪的发展趋势———手持式、偏振、微束分析等进行了评述,并对其技术的发展方向进行了展望。     

环境污染物的免疫传感检测方法进展

本文首先从免疫传感器的构建开始论述,对近5年有关免疫传感器用于环境污染物检测的文章进行了分类和归纳,对其中的三大热点进行了详细介绍,即全内反射荧光、光波导模式谱和表面等离子共振免疫传感器.其次对该领域的研究现状进行了分析,重点从信号放大技术、多组分检测、传感器的再生以及自动化和小型化等方面进行评述.最后,对免疫传感器用于环境污染物检测的发展趋势作了讨论.     

毛细管电泳免疫分析的发展及动向

毛细管电泳免疫分析是一种新兴的免疫分析技术，较常规免疫分析方法具有很多的优越性。本文对毛细管电泳免疫分析近几年的发展情况作了总结，并对其发展的动向作了评述。     

三维荧光光谱技术分析应用进展

本文对三维荧光技术的发展及其分析应用作了回顾,评述与展望,引用文献７５篇。     

时间分辨荧光免疫分析的新进展

对2003~2015年间时间分辨荧光免疫分析(TRFIA)的新进展作了综述,包括荧光共振能量转移TRFIA、酶放大TRFIA和基于纳米颗粒的TRFIA,以及时间分辨荧光免疫分析与流动注射分析、实时荧光定量聚合酶链式反应(PCR)等技术联用的应用情况,并对TRFIA的发展前景进行了展望(引用文献82篇)。     

脂质体放大免疫分析的进展

对脂质体放大免疫分析的各种测定模式、所使用的标记物及其在临床和其它分析技术中的应用作了较详细的评述，涉及参考文献５５篇。     

稀土螯合物探针及其在时间分辨荧光免疫分析中的应用

本文总结了近年来所用的稀土螯合物探针及其在时间分辨荧光免疫分析中的应用，着重介绍了稀土螯合物探针标记的原理和特点，评述了ＬＫＢ体系和ＣｙｂｅｒＦｌｕｏｒ体系的相对优缺点，展望了荧光免疫分析的发展趋势     

动物源食品中β-兴奋剂残留物检测技术的进展评述

对动物源食品中β-兴奋剂残留物的检测技术的进展作了评述。内容涉及试样处理技术以及多种用于分析的测试技术，诸如色谱、免疫分析、生物传感技术等。引述文献31篇。     

荧光偏振技术分析全血XPD基因单核苷酸多态性的研究

将荧光偏振与非对称基因扩增技术联用,建立了可用于检测全血XPD基因单核苷酸多态性的新方法。用不等量(1∶5)的XPD基因上、下游引物对含单核苷酸多态性位点的目的片段进行非对称扩增,再用两种单核苷酸多态性序列特异的荧光标记探针对扩增产物进行检测。由于扩增得到的单链片段能够与各自不同的荧光标记探针特异结合,使荧光标记分子的分子量增加,偏振值(FP)增高。通过检测增高的FP值,可确定目的片段单核苷酸多态性。采用本方法对98例全血的XPD基因第751位密码子进行了单核苷酸多态性分析,并与传统的荧光偏振检测方法进行了比较,取得满意结果。     

Stopped-flow fluorescence polarization immunoassay

A general survey of the analytical application of kinetic methodology in fluorescence polarization immunoassay (FPIA) is presented. Stopped-flow mixing technique (SF) allows the initial rate of the immunochemical reaction between the tracer and the antibody to be obtained, which is used as the analytical parameter instead of the equilibrium signal used in conventional FPIA. The instrumentation required is described and the features of the analytical methods proposed are compared with those obtained by conventional FPIA. The usefulness of SF-FPIA for routine screening in clinical, environmental and food analysis is discussed.     

A simple, rapid and high-throughput fluorescence polarization immunoassay for simultaneous detection of organophosphorus pesticides in vegetable and environmental water samples

A simple, rapid and high-throughput fluorescent polarization immunoassay (FPIA) for simultaneous determination of organophosphorus pesticides (OPs) using a broad-specificity monoclonal antibody was developed. The effects of tracer structure, tracer concentration, antibody dilution, methanol content and matrix effect on FPIA performance were studied. The FPIA can detect 5 OPs simultaneously with a limit of detection below 10 ng mL(-1). The time required for the equilibrium of antibody-antigen interaction was less than 10 min. The recovery from spiked vegetable and environmental samples ranged from 71.3% to 126.8%, with the coefficient of variations ranging from 3.5% to 14.5%. The developed FPIA was applied to samples, followed by confirmation with high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis. The developed FPIA demonstrated good accuracy and reproducibility, and is suitable for rapid and high-throughput screening for OP contamination with high-efficiency and low cost.     

Microwave and ultrasonic extraction of chlorophenoxy acids from soil and their determination by fluorescence polarization immunoassay

Procedures were developed for the ultrasonic and microwave extraction of pesticides, 2,4-dichlorophenoxyacetic (2,4-D) and 2,4,5-trichlophenoxyacetic (2,4,5-T) acids from soils for the subsequent determination by fluorescence polarization immunoassay (FPIA). The effect of the matrix composition of soils on the FPIA results was studied, and the optimum extractants and extraction conditions were selected. It was found that 40% ethanol is optimum for both extraction and FPIA determination, because it does not cause antibody denaturation. The recovery of pesticides in soil was 80–132% for 2,4-D and 101–138% for 2,4,5-T. Microwave extraction is more efficient than ultrasonic extraction for the determination of 2,4-D and 2,4,5-T in soil. The detection limit in soil and the analytical range are 2 and 4–200 μg/g, respectively, for 2,4-D and 20 and 80–5000 μg/g, respectively, for 2,4,5-T. Results of the determination of 2,4-D in soil by FPIA are in good agreement with the results of the determination by high-performance liquid chromatography. The procedures can be used for the rapid determination of chlorophenoxy acids in soils.     

A clinical trial for therapeutic drug monitoring using microchip-based fluorescence polarization immunoassay

Microchip analysis is a promising method for therapeutic drug monitoring. This led us to evaluate a microchip-based fluorescence polarization immunoassay (FPIA) system for point-of-care testing on patients being treated with theophylline. The sera were collected from 20 patients being treated with theophylline. Fluorescence polarization was measured on the microchip and theophylline concentrations in serum were obtained. Regression analysis of the correlations was done between the results given by the microchip-based FPIA and the conventional cloned enzyme donor immunoassay (CEDIA), and between the results given by the microchip-based FPIA and the conventional particle-enhanced turbidimetric inhibition immunoassay (PETINIA). We successfully carried out a quantitative analysis of theophylline in serum at values near its therapeutic range in 65 s. The results obtained by the microchip-based FPIA correlated well with CEDIA and PETINIA results; the correlation coefficients (R ²) were 0.986 and 0.989, respectively. The FPIA system is a simple and rapid method for point-of-care testing of drugs in serum, and its accuracy is the same as the conventional CEDIA and PETINIA. It is essential to use real samples from patients and to confirm good correlations with conventional methods for a study on the realization of microchip.     

荧光偏振免疫分析方法快速检测沙拉沙星残留

以异硫氰酸荧光素(FITC)标记沙拉沙星合成荧光标记物,采用薄层色谱法提纯,优化了反应时间、标记物和抗体的工作浓度,建立了沙拉沙星的快速荧光偏振免疫分析法( FPIA).本方法测定沙拉沙星在缓冲液中的半数抑制浓度(IC50)为43.2 μg/L;检测范围为5.7～327 μg/L,可以达到国家规定的动物性食品中兽药最高残留限量(80 μg/kg)的要求.本研究考察了FPIA测定沙拉沙星的动力学过程及对其它4种喹诺酮类药物的交叉反应.结果表明,环丙沙星、恩诺沙星、加替沙星及氧氟沙星的交叉反应率分别为3.3％,1.8％,1.7％和0.7％.在牛奶和猪尿中沙拉沙星的回收率分别在71％～94％和74％～102％之间.本方法操作简单快捷,整个检测过程只需5 min、而且灵敏度较高、特异性强,适用于动物性食品中沙拉沙星残留的快速筛选检测.     

Fluorescence polarization immunoassays for metal ions

Antibodies raised against a given metal ion complex of a polyaminopolycarboxylate chelating agent can display specificity for the immunizing chelate and, when used in conjunction with a fluorophore-labeled analog of that chelate, can form the basis for highly sensitive and specific methods for detecting that metal ion by competitive inhibition fluorescence polarization immunoassay (FPIA). Chelate complexes of ethylenediamine-N,N,N',N'- tetraacetic acid (EDTA) and of a hetrocyclic ring-substituted derivative of diethylenetriamine-N, N', N"-triacetic acid (DTTA) have been used to configure such assays for the heavy metal ions lead(II) and cadmium(II) respectively. Limits of detection for the 1:1 metal chelates under ideal conditions are 20 ppt for lead(II) and below 100 ppt for cadmium(II). Standard curves for 0 - 100 nM cadmium (II) chelate can be constructed in the presence of fixed 250 nM concentrations of the corresponding, potentially cross-reactive chelates of zinc(II), copper(II) and mercury(II). Cross-reactivity of the lead (II) FPIA with 15 non-target metals is below 0.2% in all cases except for mercury(II) (0.37%). These characteristics have allowed the development of FPIA methods for the quantitative analysis of lead in a variety of samples relevant to environmental monitoring, including soil, dust, solid wastes and drinking water. Although applied thus far to heavy metals that are of concern as toxic contaminants in the environment, anti-chelate FPIA methods are also in principle applicable to a wide variety of other metal ions, including precious metals and various transition and main group elements used or monitored in a range of industrial applications. As conventional methods for trace metal analysis based on atomic spectroscopy are relatively slow, expensive and cumbersome, anti-chelate FPIA methods have the potential to supplant many existing techniques and in so doing extend the use of immunoassay technology beyond the biomedical, veterinary and agricultural spheres in which it has historically found use.     

Monoclonal Antibody-Based Fluorescence Polarization Immunoassay for Furazolidone in Feed

A rapid, specific, and sensitive fluorescence polarization immunoassay (FPIA) method was developed to detect residues of furazolidone, a forbidden drug in feed. An immuno-hapten was designed for monoclonal antibody preparation. Furthermore, seven novel tracers were synthesized and the “heterology tracer” could improve the sensitivity of FPIA significantly. An optimized FPIA method was established with a cross-reactivity of less than 0.1%, IC₅₀ of 5.5 ng/mL. The limits of detection (LOD) calculated from feed samples were 0.5–0.9 ng/mL. Recoveries of fortified samples at levels of 5, 20, and 50 ng/mL ranged from 79.0% to 85.0%. The coefficients of variation were less than 12%.     

Fluorescence polarization immunoassays and related methods for simple, high-throughput screening of small molecules

Fluorescence polarization immunoassay (FPIA) is a homogeneous (without separation) competitive immunoassay method based on the increase in fluorescence polarization (FP) of fluorescent-labeled small antigens when bound by specific antibody. The minimum detectable quantity of FPIAs with fluorescein label (about 0.1 ng analyte) is comparable with chromatography and ELISA methods, although this may be limited by sample matrix interference. Because of its simplicity and speed, FPIA is readily automated and therefore suitable for high-throughput screening (HTS) in a variety of application areas. Systems that involve binding of ligands to receptor proteins are also susceptible to analysis by analogous FP methods employing fluorescent-labeled ligand and HTS applications have been developed, notably for use in candidate drug screening.     

Fluorescence polarization immunoassay for salinomycin based on monoclonal antibodies

A fluorescence polarization immunoassay(FPIA) for the determination of salinomycin(SAL) was developed by using anti-SAL monoclonal antibodies(mAb).Fluorescein labeled SAL(tracer) was synthesized by the N-hydroxysuccinimide active ester method and purified using thin layer chromatography(TLC).The developed FPIA for SAL had a dynamic range from 0.60 to 2193 ng/mL with an IC50 value of 33.2 ng/mL and a detection limit(LOD) of 0.08 ng/mL.No significant cross-reactivities were observed with other drugs but 67.6%...     

Determination of 2,4-dichlorophenoxyacetic acid in cereals by fluorescence polarization immunoassay

A fluorescence polarization immunoassay (FPIA) procedure is developed for the determination of 2,4-dichlorophenoxyacetic acid in cereal grains using a Sentry-200 portable device. We synthesized tracers, that is, antigens labeled with fluorescein derivatives, based on two fluorescent compounds and two chlorophenoxyacids. The tracer synthesized from 2,4-dichlorophenoxyacetic acid and 4-aminomethylfluorescein is found to be optimal for the FPIA of 2,4-dichlorophenoxyacetic acid. The limit of detection for 2,4-dichlorophenoxyacetic acid in flour was 40 ng/g; the analytical range is from 80 to 1000 ng/g. The percentage of recovery was 85 ± 7% at the relative standard deviation (RSD) 2–10%.