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Antibody compound for detecting human prepalin retinol binding protein 4, as well as immunochromatography test card and kit

An antibody complex and protein binding technology, applied in the field of biomedicine, to achieve the effect of strong specificity and good sensitivity

Inactive Publication Date: 2012-08-29
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, through a patent search, it was found that there is currently no relevant kit patent for the diagnosis of renal tubular injury by using colloidal gold immunochromatography to detect RBP4

Method used

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  • Antibody compound for detecting human prepalin retinol binding protein 4, as well as immunochromatography test card and kit
  • Antibody compound for detecting human prepalin retinol binding protein 4, as well as immunochromatography test card and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: the preparation of colloidal gold-RBP antibody complex

[0023] Use 0.1mol / LK 2 CO 3 Adjust the pH of 1ml of colloidal gold (particle size 20nm) solution with a concentration of 1mg / mL to 8.5, then add 5μg of RBP4 monoclonal antibody 1 to the colloidal gold solution, shake and react at room temperature for 10 minutes, and then block with 10% BSA solution , centrifuged at low temperature, resuspended to the original volume with boric acid buffer solution with a concentration of 0.002 mol / L, pH=9.0, purified by centrifugation again, discarded the supernatant, and stored the lower layer at 4°C.

Embodiment 2

[0024] Example 2: Preparation of Retinol Binding Protein 4 (RBP4) Immunochromatographic Detection Card

[0025] 2.1 Preparation of sample pad: choose glass cellulose membrane as the sample pad material, put it into the treatment solution (pH = 7.4, 0.01M phosphate buffer PBS, containing 0.5% ~ 1.0% BSA, 0.3% ~ 0.8% vinylpyrrolidone PVP, 0.1% ~ 0.3% Tween-20) soaked in 10 minutes, took out, and dried at 37 ℃ for later use.

[0026] 2.2 Preparation of conjugation pads: select glass cellulose membrane as the conjugation pad material, and redisperse the colloidal gold-RBP antibody complex prepared in the above example 1 in colloidal gold diluent (0.05mol / L , pH=8.0 Tris-HCl buffer solution, containing 0.5%~1.0%BSA, 0.3%~0.8% polyvinylpyrrolidone, 0.2%~0.5% polyvinyl alcohol 20,000, 1.0%~3.0% sucrose, 0.1%~0.3 % Tween-20, all by weight), and sprayed on the bonding pad, and dried at 37°C.

[0027] 2.3 Coating of nitrocellulose membrane (NC membrane) and preparation of analytical m...

Embodiment 3

[0029] Embodiment 3: the preparation of sample diluent

[0030] Prepare a certain concentration of phosphate buffer solution PBS (0.01mol / L, pH7.2-7.4), add an appropriate amount of protective protein BSA (final concentration 5%), and finally add preservative Proclin 300 (0.3%-0.5%). Dispense the prepared sample diluent into 15mL plastic reagent bottles to obtain the RBP4 sample diluent.

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Abstract

The invention relates to the field of biomedicine and particularly relates to an antibody compound for detecting human prepalin retinol binding protein 4, as well as an immunochromatography test card. A monoclonal antibody 1 of human prepalin retinol binding protein 4 is connected with colloidal gold granules, and then the antibody compound is obtained and used for preparing the immunochromatography test card for human prepalin retinol binding protein 4, and a kit. The invention has the advantages of good sensitivity, strong specificity and the like.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to an antibody complex for detecting human retinol binding protein 4 and an immunochromatographic detection card. Background technique [0002] Retinol Binding Protein (RBP) is a kind of carrier protein responsible for transporting retinol from the liver to various tissue cells in the human body, and plays an important role in the physiological function of retinol. RBP is mainly synthesized in the liver. The RBP-retinol polymer secreted in the liver forms a stable protein complex with transferrin (Transthyretin, TTR) and circulates in the blood system. The RBP-retinol-TTR complex disintegrates after the release of retinol, and RBP exists in the blood system in a free form, is filtered by the glomerulus, and then reabsorbed and degraded by the proximal tubular epithelial cells. Therefore, normal human urine hardly contains RBP (the content is less than 300ng / ml). If the kidney tubule is...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/558
Inventor 邢国杰沈鹤柏
Owner SHANGHAI NORMAL UNIVERSITY
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