Enzymes chemiluminescent detection

Blankenstein G., Preuschoff F., Spohn U., Mohr K.H., Kula M.R., Determination of L-glutamate and L-glutamine by flow-injection analysis and chemiluminescence detection comparison of an enzyme column and enzyme membrane sensor, Anal. Chim. Acta 1993 271 231-237. 

J. Yakovleva, R. Davidsson, M. Bengtsson, T. Laurell, and J. Emneus, Microfluidic enzyme immunosensors with immobilised protein A and G using chemiluminescence detection. Biosens. Bioelectron. 19, 21-34 (2003). 

Therefore, this sensor integrates a biochemical and a chemical reaction with a prior separation (dialysis) and chemiluminescence detection. The process involves the following steps (a) dialysis of the enzyme (6) enzymatic oxidation of the reagent (c) derivatization of hydrogen peroxide and d) detection of the chemiluminescence produced. Such an original approach offers several advantages over similar methodologies, namely ... 

We also examined the preservation stability of the photoimmobilized antibodies. Although the reactivity of the antibodies dropped away over a period of 10 days when they were stored at room temperature, it was maintained for about 2 months when stored at 4°C. This result is acceptable in terms of commercial viability, though further increase in stability would be preferable. We next examined the sensitivity for the immunochips. An immu-nochip usually has a two-dimensional surface, so the detection limit for antigens can be estimated from the amount of immobilized antibodies that are present. It was difficult to increase the sensitivity of a detection system which uses a photoluminescence probe. However, we succeeded in obtaining higher sensitivity for an immunochip in which we adopted a chemiluminescence detection system using an enzyme reaction. 

Directly labeled chemiluminescent systems produce <1 photon/label and require complex chemical synthesis to produce each new labeled molecule (1,2). In contrast, chemiluminescent detection of enzyme labels combines the advantages of a high specific activity label with the convenience of relatively simple coupling chemistries, which use commercial reagents. A number of enzyme labels can be detected via chemiluminescent or bioluminescent reac-... 

Surugiu et al. [67] have introduced an Enzyme Immuno-Like Assays (EzILA) for the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). The label was a 2,4-D conjugate with the tobacco peroxidase (TOP) enzyme, which allows for both colorimetric and chemiluminescent detection. In this case, the polymer imprinted with 2,4-D was synthesized in the form of microspheres. In contrast, despite its higher binding capacity for radiolabeled 2,4-D, a conventional MIP prepared by bulk polymerization showed only weak binding of the 2,4-D-TOP tracer. 

Yakovleva, J., Davidsson, R., Lobanova, A., Bengtsson, M., Eremin, S., Laurell, T., Emneus, J., Microfluidic enzyme immunoassay using silicon microchip with immobilized antibodies and chemiluminescence detection. Anal. Chem. 2002, 74(13), 2994-3004. 

Rong, W., Kutter, J.P., On-line chemiluminescence detection of bioprocesses using polymer-based microchips with immobilized enzymes. Micro Total Analysis Systems, Proceedings 5th pTAS Symposium, Monterey, CA, Oct. 21-25, 2001, 181-182. 

Fan and Zhang determined acetylcholine and choline in rat brain tissue by a fluorescence immunoassay method, making use of immobilized enzymes and chemiluminescence detection [50]. Tissue was homogenized with a 10 fold volume of 0.6 M HC104, the homogenates were kept on ice for 30 min, and then centrifuged at 2000 G for 20 min. The pellets were... 

Li HA, Cao ZJ, Zhang YH, Lau CW, Lu JZ (2010) Combination of quantum dot fluorescence with enzyme chemiluminescence for multiplexed detection of lung cancer biomarkers. Anal Meth 2 1236-1242... 

More recently, automated immunoanalyzer methods for DPD have become commercially available. The most widely used is a solid-phase, competitive EIA with chemiluminescence detection. The solid-phase antibody is incubated with serum or calibrator and ALP conjugated to DPD. After washing, the antibody-antigen-enzyme complexes are determined after the addition of substrate,... 

A heterogeneous solid-phase enzyme immunoassay for DHEA-S is commercially available. This method uses horseradish peroxidase as enzyme label, rabbit anti-DHEA-S-coated microtiter wells, and tetramethylbenzidine as substrate. The assay is stated to have a lower limit of detection of 15ng/dL (40.6nmol/L). An automated method employing chemiluminescent detection of alkaline phosphatase-labeled conjugate has been described. This assay reported a lower limit of detection of 2pg/mL (54.0nmol/L). 

Dose-response curves obtained with the chemiluminescence detection of enzyme activity under the optimal conditions, are shown in Figure 3. The linear range is... 

EHJM Jansen, et al. Comparison between spectrophotometric and chemiluminescence detection in enzyme immunoassays for nortestosterone. Anal Chim 227 109,... 

Although it is not the purpose of this article to champion one technology over another, it is fair to say that chemiluminescence offers the sensitivity of fluorescence detection without some of the attendant problems of luminescent emission from analytical samples. Thus, chemiluminescence detection can be comparable to and, with the aid of enzyme amplification, even superior to that of I. This is not to say that there are no problems associated with chemiluminescence-based analytical techniques. As we shall see later (Section 2.5), the actual signal from chemiluminescent molecules is, in most cases, a transient flash, lasting... 

A second example is the determination of glucose, free cholesterol, creatinine, and lactic acid (figure 21.8). This method uses on-line immobilized enzyme reactors followed by chemiluminescence detection of the hydrogen peroxide enzyme reaction product reaction with luminol. Detection limits of 0.1 mg dl were observed. 

Mickova et al. (2004) have described an enzyme-linked immunosorbent assays (ELISA) with chemiluminescent detection for the determination of carbaryl, carbo-furan and methiocarb. These compounds can also be detected by colorimetric-ELISA methods. The authors have reported very low detection levels of 0.03, 0.007 and... 

Xu Q, Zhu JJ, Hu XY (2007) Ordered mesoporous polyanifine film as a new matrix for enzyme immobilization and biosensor construction. Anal Chun Acta 597(1) 151—156 Yakovleva J, Davidsson R, Lobanova A et ak (2002) Enzyme immunoassay using silicon microchip with immobilized antibodies and chemiluminescence detection. Antd Chem 74(13) 2994—3004 Yamada H, Kobayashi M (1996) Nitrile hydiatase and its application to industrial production of acrylamide. Biosci Biotechnol Biochem 60 1391—1400 Yamamoto K, Ohgam T, Torimura M et al. (2000) Highly-sensitive flow injection determination of hydrogen peroxide with a peroxidase-immobilized electrode and its application to clinical chemistry. Anal Chim Acta 40(2) 201—207... 

---