Alkaline phosphatase assay automation

Part—I has three chapters that exclusively deal with General Aspects of pharmaceutical analysis. Chapter 1 focuses on the pharmaceutical chemicals and their respective purity and management. Critical information with regard to description of the finished product, sampling procedures, bioavailability, identification tests, physical constants and miscellaneous characteristics, such as ash values, loss on drying, clarity and color of solution, specific tests, limit tests of metallic and non-metallic impurities, limits of moisture content, volatile and non-volatile matter and lastly residue on ignition have also been dealt with. Each section provides adequate procedural details supported by ample typical examples from the Official Compendia. Chapter 2 embraces the theory and technique of quantitative analysis with specific emphasis on volumetric analysis, volumetric apparatus, their specifications, standardization and utility. It also includes biomedical analytical chemistry, colorimetric assays, theory and assay of biochemicals, such as urea, bilirubin, cholesterol and enzymatic assays, such as alkaline phosphatase, lactate dehydrogenase, salient features of radioimmunoassay and automated methods of chemical analysis. Chapter 3 provides special emphasis on errors in pharmaceutical analysis and their statistical validation. The first aspect is related to errors in pharmaceutical analysis and embodies classification of errors, accuracy, precision and makes... 

ST53 Butch, A.W., Goodnow, T.T., Brown, W.S., McCellan, A., Kessler, G. and Scott, M.G. (1989). Stratus automated creatine kinase-MB assay evaluated Identification and elimination of falsely increased results associated with a high-molecular-mass form of alkaline phosphatase. Clin. Chem. 35, 2048-2053. 

Chemiluminescence assays are ultrasensitive (attomole to zeptomole detection limits) and have wide dynamic ranges. They are now widely used in automated immunoassay and DNA probe assay systems, (e.g., acridinium ester and acri-dinium sulfonamide labels and 1,2-dioxetane substrates for alkaline phosphatase labels and the enhanced-luminol reaction for horseradish peroxidase labels [see Chapter 9]). 

Immunometric assays for TSH are available commercially as manual kit procedures or for use on automated systems. For practical reasons, nonisotopic methods dominate the market and have replaced radioactive tracer methods in most routine laboratories. The majority of immunometric TSH assays label the detection antibody with chemiluminescent labelled molecules other labels include peroxidase or alkaline phosphatase and sensitive photo-metric and fluorescenri molecules. Other assays are based on the use of fluorescent labels using europium chelates chemiluminescent compounds such as acri-dinium esters or ruthenium or bioluminescent molecules such as recombinant aequorin. ... 

Considerable effort was expended in the development of alternative technologies that did not require the use and measurement of radioactivity. A number of nonisotopic assays for T4 were subsequently developed commercially for use on fuUy automated immunoassay systems or for use with existing chemistry analyzers. According to a 2002 College of American Pathologists Ligand Assay Survey, more than 95% of laboratories now use a nonisotopic T4 method. A variety of different labels were used to construct these nonisotopic assays. Enzymes such as horseradish peroxidase, alkaline phosphatase, and [3-D-galactosidase were the most... 

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). 

A number of nonisotopic immunoassays for estradiol have been developed and adapted for use on fuHy automated immunoassay systems. All are heterogeneous assays (separation step needed), but most are direct assays and do not require prehminary extraction. Most procedures offer the convenience of solid-phase separation methods. For routine clinical applications, the greatest experience is with enzyme immunoassays. Most commercial enzyme immunoassays use horseradish peroxidase or alkaline phosphatase to label estradiol antigens enzyme activity is determined using a variety of photometric,fluorescent,or chemiluminescent substrates. ... 

Hfll HD, Summer GK, Waters MD (1968) An automated fluorometric assay for alkaline phosphatase using 3-O-methylfluorescein phosphate. Anal Biochem 24 9—17... 

Key words High-throughput screens, HTS, Chemical libraries. Laboratory automation. Automated assays. Robotics, Phosphatase assays. Assay development. Implementation, Enzymes, Alkaline phosphatases. Tissue-nonspecific alkaline phosphatase (TNAP)... 

Fig. 4. Schematic diagram of the steps in the automated PCR/OLA procedure performed with a robotic workstation. The assay contains three steps (1) DNA target amplification (2) analysis of target nucleotide sequences with biotin (B)-labeled and digoxigenin (D)-labeled oligonucleotide probes and T4 DNA ligase (L) and (3) capture of the biotin-labeled probes on streptavidin (SA)-coated microtiter wells and analysis for covalently linked digoxigenin by using an ELISA procedure with alkaline phosphatase (AP)-conjugated antidigoxigenin (aD) antibodies and a substrate (S). Reprinted with the permission of Nickerson el al. (N2) and the Proc. Natl. Acad. Sci. (U.SA.).

To solve the problem, we have developed a highly specific and sensitive assay for C-peptide in serum and plasma using specific monoclonal antibody (MoAb) to the N-terminal of the C-peptide molecule. In this report, we describe the assay performance of C-peptide on the LUMIPULSE system. The system is a fully automated chemiluminescent enzyme immunoassay (CLEIA) system that uses AMPPD as a substrate for alkaline phosphatase and ferrite micro-particles as a solid phase. ... 

Beilstein Handbook Reference) AI3-03947 Aminomethyl propanol Aminomethylprop-anol AMP AMP 75 AMP 95 AMP Regular BRN 0506979 Caswell No. 037 Corrguard 75 EINECS 204-709-8 EPA Pesticide Chemical Code 005801 HSDB 5606 Hydroxy-tert-butylamine lsobutanol-2-amine KV 5088 NSC 441. Boiler water treatment ohemioal, corrosion inhibitor, carbon dioxide absorber. Widely used as a buffer and phosphate acceptor in assay of phosphatases. Suitable as buffer for manual and automated determination of alkaline phosphatase using 4-nitrophenyl phosphate as substrate. Solid mp = 25.5° bp = 165.5° d O = 0.934 soluble in CCI4, freely soluble in H2O LDso (rat orl) = 2900 mg/kg. Lancaster Synthesis Co. Lancaster Synthesis Ltd. 

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