Glycated albumin detection immunochromatography test trip and preparation method thereof
An immunochromatographic test paper and glycosylated albumin technology, which is applied in the field of immunoassay, can solve the problems of unsuitable clinical routine development, high cost, and small sample size, and achieve easy large-scale production, high sensitivity, and simple preparation method Effect
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[0038] A method for preparing an immunochromatographic test strip for detecting glycated albumin, comprising the following steps:
[0039] 1) Preparation of phenylboronic acid markers: Dilute the fluorescent material with buffer, add phenylboronic acid solution, react at 20-25°C for at least 1 hour, use G25 gel column for separation and purification, and collect markers. That is, phenylboronic acid marker;
[0040] 2) Preparation of the sample pad: use cellulose membrane as the solid phase material of the sample pad, soak it in the first phosphate buffer solution with a substance concentration of 0.01-0.3mol / L, and the pH value of the first phosphate buffer solution is 7.2- 7.6, after soaking, dry it to get the sample pad;
[0041] 3) Preparation of the conjugate pad: use glass cellulose membrane as the solid phase material of the conjugate pad, dilute the phenylboronic acid marker with the second phosphate buffer with a concentration of 0.01-0.1mol / L and a pH value of 7.2 ,...
Embodiment 1
[0051] A method for preparing an immunochromatographic test strip for detecting glycated albumin, comprising the following steps:
[0052] 1) Preparation of phenylboronic acid markers:
[0053] Dilute anti-albumin monoclonal antibody and goat anti-mouse IgG antibody to 1mg / ml with sodium bicarbonate-sodium carbonate solution with concentration of 0.1mol / L and pH of 9.6 respectively, take 5ml of antibody solution, respectively Add 30 mg fluorescent material metalloporphyrin solution, stir well, incubate at room temperature for 1 hour, and mix every 15 minutes. Finally, use a G25 gel column for separation and purification to collect the labeled metalloporphyrin-labeled anti-albumin antibody and goat anti-mouse IgG antibody. The first phosphate buffer solution dilution, wherein the first phosphate buffer solution includes polyethylene glycol containing 0.1% by mass percentage, 2.0% bovine serum albumin, and 0.05% first surfactant, sealed with a reagent bottle , Stored at 2-8°C....
Embodiment 2
[0069] Same as Example 1, the difference is:
[0070]In step 1 and step 2, buffer solution is identical with the first phosphate buffer saline component, and its substance concentration is 0.05mol / L, and pH value is 7.4, and it comprises: by mass percentage, 0.01% polyethylene glycol Alcohol, 1% bovine serum albumin and 0.01% primary surfactant.
[0071] In step 3, the second phosphate buffer solution has a substance concentration of 0.5mol / L and a pH value of 7.2, which includes: in terms of mass percentage, 1% bovine serum albumin, 0.1% polyethylene glycol, 0.5% sucrose and 0.01% secondary surfactant.
[0072] The first surfactant and the second surfactant were Triton X-100.
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