Method for producing solid element for detection of multiple analytes and its use, and system comprising said element
An analyte and solid-state element technology, applied in the field of components and devices for simultaneous determination of multiple analytes, can solve the problems of time-consuming and expensive
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example 1
[0101] example 1 Sulfa determination
[0102] In this example, 12 antibodies, all specific for a single sulfonamide, were covalently immobilized by contact interactions on discrete areas of a flat ceramic (alumina) substrate with a chemically modified surface. The determination of multiple analytes is accomplished using a competitive immunoassay.
[0103] In more detail, ceramic substrates (1 cm × 1 cm) were ultrasonically cleaned with alkaline detergent (RBS35, 5% v / v) followed by two times deionized water, and then placed in 6M HCl for 16 h . The tablets were then placed in chromic acid in an ultrasonic bath for 1 hour.
[0104] The substrate was thoroughly rinsed with double deionized water and acetone, and then placed in an oven at 120°C for two hours to dry. After this pretreatment, the organic silane Y-glycidyloxytrimethoxysilane (10% V / V) in anhydrous toluene, 4-dimethylaminopyridine (1.25g / L) and triethylamine (1 % V / V) to silanize the above substrate. This mixt...
example 2
[0109] Example 2 Hormone determination
[0110] In this example, a multi-analyte assay was performed on three large molecular weight hormones, namely prolactin (PL), follicle stimulating hormone (FSH), and luteinizing hormone (LH). This example shows the determination of multiple analytes for a Sandwich-based immunoassay. When the above three hormones were measured in the same group, no significant cross-reactivity was seen.
[0111] The chemical pretreatment and silanization procedures were exactly as described in Example 1. PL, FSH or LH monoclonal antibodies (approximately 20 nl of antibody dispensed) were immobilized on discrete areas of the chemically modified substrate. The determination of various analytes was performed on both silicon and ceramic substrates with epoxy surfaces as described in Example 1.
[0112] For the assay, 150 μl of multi-LH / PL / FSH serum-based standard and 150 μl of diluted assay buffer were added to the above components and incubated at room ...
example 3
[0113] Example 3 Sulfa determination
[0114] In contrast to Example 1, multiple sulfonamide assays were performed using microchannels. The components are shown in Figure 11. In this example, the reagent addition container 21 is 2 mm x 2 mm, 300 μm deep (volume 1.2 μl), the channels 23 are each 5 mm long, 200 μm wide, and 100 μm deep (volume 100 nl), and the container 24 is 1.9 mm x 8.6 mm, 300 μm Deep (volume 4.9 μl).
[0115] The surface was chemically modified as described in Example 1. Antibodies were added to each channel and incubated at 37°C for 2 hours. The substrate was then protected and washed as in Example 1.
[0116] Various sulfa standards (200 μl) and sulfa-horseradish peroxidase conjugate (100 μl) were mixed. 1 µl of the resulting reagent was pipetted into each of the above containers to feed the antibody-coated channels for each sulfonamide. Suitable samples contain 10 or 100 ng / ml of all sulfonamides.
[0117] Reagent flow is achieved by capillary ac...
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