An antibody chip kit for early diagnosis of acute kidney injury

An acute kidney injury and antibody chip technology, applied in the field of biomedicine, can solve the problems of single detection index and complicated operation, and achieve the effects of non-cross-contamination, uniform distribution, and increased stability.

Active Publication Date: 2016-06-01
RAYBIOTECH INC GUANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The kit of the present invention can detect 20 specific early diagnostic indicators of acute kidney injury, overcomes the defects of the prior art such as cumbersome operation, single detection indicator, and low sensitivity, and is cheap, convenient, sensitive, accurate, high-throughput, The advantages of small amount of specimen, can be promoted and scaled up in ordinary laboratories, etc.

Method used

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  • An antibody chip kit for early diagnosis of acute kidney injury
  • An antibody chip kit for early diagnosis of acute kidney injury
  • An antibody chip kit for early diagnosis of acute kidney injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Determination of antibody chip spotting conditions.

[0025] 1. Spotting concentration of the capture antibody: put the capture antibody in different buffers (sterile water, PBS buffer, PBS containing different concentrations of bovine albumin, PBS buffer containing different concentrations of glycerol, etc.) at a concentration of 2x Dilute it, spot it on the surface of the chip with a non-contact spotter, and compare its activity and stability with a sandwich ELISA method. Experiments show that diluting the capture antibody with PBS buffer containing 0.01-10g / 100ml bovine albumin has the best linear rate and stability.

[0026] 2. Condition control of spotting: when the capture antibody is directly spotted on the surface of the slide at room temperature, the spot of the capture antibody is likely to produce hollow spots, and the activity of different antibody spots is quite different, and it is often found that there will be some spots in the final generated...

Embodiment 2

[0027] Example 2: Preparation of the antibody chip kit for early diagnosis of acute kidney injury according to the present invention.

[0028] In order to detect whether there are corresponding acute kidney injury markers in the sample, prepare slides immobilized with specific antibodies against the following acute kidney injury markers: kidney injury molecule 1, albumin, osteopontin, trefoil factor 3, β2-microglobulin, clusterin, chemokine ligand 16, non-metastatic melanoma glycoprotein B, hepatic fatty acid binding protein, monocyte chemoattractant protein-1, soluble tumor necrosis factor receptor 1, calbindin -1, γ-interferon-inducible protein 10, cystatin C, hepatocyte growth factor, macrophage migration inhibitory factor, neutrophil gelatinase-associated lipocalin, tissue inhibitor of matrix metalloenzymes-1, vascular cells Adhesin 1, vascular endothelial growth factor.

[0029] 1. The source of the antibody:

[0030] Specific antibodies against the proteins listed in T...

Embodiment 3

[0043] Example 3: An experiment of quantitatively detecting markers of acute kidney injury using the kit of the present invention.

[0044] 1. Complete drying of the slide chip

[0045] Take the slide chip out of the box, and after equilibrating at room temperature for 20-30 minutes, open the packaging bag, peel off the sealing strip, and then place the chip in a vacuum desiccator or dry at room temperature for 1-2 hours.

[0046] 2. Gradient dilution of acute kidney injury marker standard

[0047] 2.1 Add 500 μl of sample diluent to the vial of acute kidney injury marker standard mixture, and redissolve the standard. Before opening the vial, give it a quick centrifuge and gently pipet up and down to dissolve the powder. Label this vial as Std1.

[0048] 2.2 Mark six clean centrifuge tubes as Std2, Std3 to Std7 respectively, and add 200 μl of sample diluent to each small tube.

[0049] 2.3 Take 100 μl of Std1 and add it to Std2 and mix gently, then take 100 μl from Std2 and...

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Abstract

The invention relates to an antibody chip kit for early diagnosis of acute kidney injury. The kit comprises an antibody chip, an acute kidney injury marker standard substance mixture, a biotin labeled acute kidney injury marker standard substance detection antibody mixture and fluorescein Cy3 labeled streptavidin, wherein the antibody chip comprises a standard structure slide and specific antibodies of 20 acute kidney injury markers fixed on the surface of the slide, three positive control; the acute kidney injury marker standard substance detection antibody mixture is a freeze-drying mixture which is prepared by mixing the 20 acute kidney injury marker standard substances at a certain dose. The kit provided by the invention can be used for detecting 20 clinically common acute kidney injury markers, overcomes the defects in the prior art of tedious operation, single detection index, low sensitivity and the like and has the advantages that the kit is cheap, convenient, sensitive, accurate, high in throughput and few in use level of specimens and can be popularized and expanded on a common laboratory and the like.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an antibody chip kit for early diagnosis of acute kidney injury. Background technique [0002] Acute kidney injury (Acute Kidney Injury, AKI), formerly known as acute renal failure (acute renal failure, ARF) referred to as acute renal failure, is a clinical critical illness. The disease is an acute kidney injury caused by a variety of etiologies. It can cause a sharp decline in the regulatory function of the nephron within a few hours to a few days, so that it cannot maintain the balance of body fluid electrolytes and excrete metabolites, resulting in hyperkalemia, metabolic acidity, etc. Poisoning and acute uremic syndrome, this syndrome is clinically known as acute renal failure. Its main pathological change is renal tubular necrosis; however, the specific pathogenesis of acute tubular necrosis has not been fully elucidated. [0003] Although the research on the pathophys...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/533G01N21/64
CPCG01N21/6402G01N33/533G01N33/6893G01N2800/347
Inventor 黄若磐鲍伟胜陈蕴如陈巧林
Owner RAYBIOTECH INC GUANGZHOU
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