免疫亲和毛细管电泳的研究进展

免疫亲和毛细管电泳方法结合了免疫分析的高特异性和毛细管电泳分离的高效、快速、样品用量少等优点,是复杂样品中特定组分分析的重要方法之一。激光诱导荧光检测器的使用以及毛细管电泳分离前免疫预富集过程的引入,可以进一步提高分析测定的灵敏度,使其能够用于痕量物质的高灵敏测定。本文结合作者所在课题组的工作,对免疫亲和毛细管电泳的两种主要模式,即均相的毛细管电泳免疫分析(CEIA)和非均相的免疫亲和毛细管电泳(IACE)的研究进展进行了综述。     

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。     

毛细管电泳免疫化学发光检测鼠血管平滑肌细胞中zmol骨形成蛋白-2

近年来,结合毛细管电泳的免疫分析研究在不断加强.特别是毛细管电泳免疫激光诱导荧光(CEIA-LIF)检测由于具有较高的灵敏度而十分引人注目.常文保等用CEIA-LIF检测雌三醇,检出限为31.6ng/L,可用于血清和尿样分析.Kennedy等用芯片CEIA-LIF检测了鼠胰腺细胞中的胰岛素,     

高效毛细管电泳在兴奋剂检测中的应用进展

对高效毛细管电泳(HPCE)在兴奋剂检测方面的工作进行了评述,讨论了高效毛细管电泳不同分离模式在兴奋剂检测中的应用,并对这一领域的发展趋势进行了展望。     

毛细管电泳免疫分析法分析雌三醇

利用一种温敏水凝胶作为毛细管电泳的填充介质，建立了毛细管电泳免疫分析血清中雌三醇的方法。研究了缓冲溶液的浓度和pH值、水凝胶的浓度、电压等因素对分析结果的影响。雌三醇的检出限和线性范围分别为31．6ng／L和50-5000ng／L。     

毛细管电泳法进行化妆品中砷的形态分析

从样品前处理、毛细管电泳修饰、进样方式、分离模式和检测条件等方面对毛细管电泳法研究化妆品中砷的形态进行讨论。在不同pH值的缓冲溶液中,用毛细管电泳法进行化妆品中砷的形态分析,测定波长为197nm。采用磷酸盐缓冲溶液,化妆品中As(Ⅲ)、二甲基胂(DMA)、对氨苯基胂酸(ANA)、一甲基胂(MMA)和As(V)等5种形态的砷可通过毛细管电泳法得到有效分离。     

毛细管电泳应用于食品中农药残留分析的晚近进展

对2000-2008年毛细管电泳在食品农药残留分析中的应用进展进行综述,共分为三部分:第一部分介绍了毛细管电泳在农药分析中的一些主要分离模式和检测方法;第二部分介绍了与毛细管电泳联用的主要样品预处理方法;第三部分评述了近年来应用于食品中农药残留分析的各种检测方法和毛细管电泳分离模式.并探讨了这一研究领域的未来发展趋势.引用文献56篇.     

人体血液蛋白的毛细管电泳分析进展

对近年来对毛细管电泳在血清蛋白、免疫蛋白、同工酶、脂蛋白、等人 血液蛋白检测上的应用和发展前景进行了综核实 。毛细管电泳以其高效，等特点很有可能成为未来临床实验室的一种不可缺少的分析技术。     

微流控芯片毛细管电泳在蛋白质分离分析中的应用研究进展

重点介绍了近年来国内外在微流控芯片毛细管电泳法用于蛋白质分离分析方面的研究进展。按照分离模式的不同,综述了各种应用于蛋白质分离的微流控芯片毛细管电泳系统,讨论了抑制芯片中的蛋白吸附的各种方法,并展望了芯片毛细管电泳系统在蛋白质分离领域的发展前景。引用文献47篇。     

亲和毛细管电泳技术及其应用

对近几年新发展起来的亲和毛细管电泳技术（ACE）的原理、分类及方法作了简要介绍,着重介绍了亲和毛细管区带电泳、毛细管亲和凝胶电泳、胶束电动色谱中的亲和电泳、亲和毛细管等电聚焦、亲和探针毛细管电泳等过程和方法。对ACE在分子生物学、生物化学中的应用及该技术在亲和常数测定、核酸片段识别、竞争免疫分析、药物先导化合物的筛选等方面的应用也作了介绍。     

Affinity protection chromatography for efficient labeling of antibodies for use in affinity capillary electrophoresis

Capillary electrophoresis immunoassay (CEIA) is shown to be substantially more sensitive to the antibody (Ab) reagent quality than are immunosorbent methods such as enzyme-linked immunosorbent assays (ELISA). Cyanine 5 (Cy5)-labeled monoclonal anti-ovalbumin (mAb*) was inactive for CEIA of ovalbumin (Ov), yet was functional in ELISA for Ov. ELISA showed the mAb* was at least ten times less active, accounting for the poor CEIA performance. Labeled polyclonal Ab was inactive for a dye to protein ratio greater than 1.6. An affinity protection chromatography procedure (APC) was developed for Ab labeling, which avoided degradation of the Ab binding site. Ov was covalently bound to cyanogen bromide activated cellulose gel in a column, and used to capture the Ab. The coupling efficiency for Ov to the gel was 74-97%, Ab could then be bound with 95-100% efficiency, and Ab* was recovered in 50% yield following labeling on the column. This procedure was performed successfully in three different laboratories, indicating the robustness of the optimized APC synthetic method. No inactive Ab* could be detected in the APC product. The CEIA detection limit for ovalbumin using APC labeled mAb was 173 nM, when [Ab*] was fixed at 163 nM. The association constants of mAb and mAb* were determined by CEIA.     

Detection of recombinant hirudin using capillary electrophoresis with laser-induced fluorescence detection

Hirudin, a thrombin inhibitor, is a polypeptide of 65 amino acids. To check purity levels and perform pharmacokinetic studies of recombinant hirudin (r-hirudin), a specific and reproducible analysis method is required. Capillary electrophoresis (CE) is rapidly becoming an important procedure for the analysis of biological molecules. Recently, CE combined with immunoassay has emerged as a new analytical technique. CE-based immunoassay (CEIA) is a sensitive and specific method combining laser-induced fluorescence (LIF) and immunoassay. Therefore, in this study, we specifically investigated fluorescence labeling and determination of r-hirudin by CEIA with a LIF detector using labeled r-hirudin and polyclonal antibody. r-Hirudin was labeled with fluorescein isothiocyanate (FITC). FITC-labeled r-hirudin was purified using high-performance liquid chromatography (HPLC). The method is based on preincubation of r-hirudin and antibody for 20 min, followed by CE analysis using an uncoated capillary. Free and bound r-hirudin were separated within 5 min using CE with high reproducibility. This study demonstrated that the CEIA method could be applied to quantitative analysis of r-hirudin in biological fluids.     

Rapid determination of multidrug resistance-associated protein in cancer cells by capillary electrophoresis immunoassay

The adenosine triphosphate (ATP) binding-cassette (ABC) transporters are a superfamily of cellular proteins that have been partly implicated as a cause of multidrug resistance (MDR) in cancer cells. The ABC superfamily consists of P-glycoprotein, multidrug resistance-associated proteins (MRP) and breast cancer-related proteins, of which MRP is of particular interest because of its ability to efflux a broader range of substrates. Since MRP1 is the most prominent member of the MRP family, a simple technique is needed for its quantification. We developed a simple, fast (total analysis time of 3h) capillary electrophoresis immunoassay (CEIA) for the quantification of MRP1 in cancer cells. MRP1 antibody was labeled with fluorescein isothiocyanate. The labeled antibody was incubated with the cell lysate for a fixed interval (1h), after which the cell lysate mixture was directly injected into the capillary to separate the complex of MRP1 and its antibody from free antibody. The noncompetitive CEIA method had a limit of detection of 0.2 nM and a good linear range (1.7-14.9×10(4) cells), and was fairly reproducible (RSD<10%). The results showed that two cell lines, A549 and RDES, expressed MRP1 in the absence of doxorubicin (DOX), with A549 registering a higher expression. Compared to DOX-free cancer cells, there was an acceleration of MRP1 expression during the 12h-exposure to DOX, after which the level of expression remained nearly constant as the intracellular accumulation of DOX decreased. The results obtained in this work indicate that the developed CEIA method is useful for relative quantification of MRPs in cancer cells.     

Combination of flow injection with electrophoresis using capillaries and chips

The technique of combined flow injection CE (FI-CE) integrates the essential favorable merits of FI and CE and can significantly expand the application of CE by utilizing the various on-line sample pretreatments and preconcentration of FI. The basic principles, instrumental developments, and applications of the FI-CE system from 2004 to 2006 are reviewed. The recent developments and applications of FI-CE are outlined.     

14-MeV neutron activation analysis: Methodology,applications, and potential

Accelerator-based 14-MeV or fast neutron activation analysis (FNAA) is a mature tecnique and few major advances in instrumentation and methodology can be expected. However, applications of the technique are numerous and continue to increase. In this paper, recent technique developments and applications of FNAA are reviewed and speculations concerning future progress in the field are presented.     

Near-infrared spectroscopy applications in pharmaceutical analysis

Near-infrared (NIR) spectroscopy is a fast and non-destructive analytical technique that offers many advantages for a broad range of industrial applications. In this work, we reviewed recent developments in the pharmaceutical domain where it can be applied from raw material identification to final product release. The characteristics of NIR allow the technique to be implemented as a process analytical technology (PAT). Moreover, recent instrumental developments open the perspectives of numerous applications in the NIR imaging area. After “Introduction”, according to their subject, the applications are discussed in the parts “Identification”, “Water content”, “Assay” and “Other applications”.     

From fundamentals to applications: recent developments in atmospheric pressure photoionization mass spectrometry

Only five years after the first publication on atmospheric pressure photoionization (APPI), this technique has evolved rapidly as a very useful complement to established ionization techniques for liquid chromatography/mass spectrometry (LC/MS). This is reflected in a rapidly increasing number of publications in this field. On the one hand, thorough studies into the photoionization mechanism have provided deep insights into the roles and influences of the solvent, the dopant and other additives. On the other hand, a large number of new and attractive applications have recently been introduced. New instrumental developments have resulted in combined APPI/ESI (PAESI) and APPI/APCI sources and a microfabricated APPI source. In this review, the most important developments within the field are summarized, focusing in particular on the applications of the technique.     

Recent applications of the ring-oven technique : A brief review

The ring-oven technique is a special type of spot analysis. In this review, papers published since 1976 are discussed. The aspects dealt with include developments in the technique and its use, the identification and determination of inorganic ions, organic substances, enzymatic analysis, applications to air and water samples, and combinations with other techniques.