Antibody chip kit for diagnosis of various tumors

A technology of antibody chips and kits, applied in the field of biomedicine, can solve the problems of cumbersome operation, single detection index, low sensitivity, etc., and achieve the effect of optimized dot matrix, uniform distribution, and easy operation

Active Publication Date: 2014-06-18
RAYBIOTECH INC GUANGZHOU +1
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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 16 tumor markers commonly used in clinical practice, overcomes the defects of the prior art such as cumbersome operation, single detection index, and low sensitivity, and is cheap, convenient, sensitive, accurate, high-throughput, and The advantages of less dosage, can be popularized and scaled in ordinary laboratories, etc.

Method used

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  • Antibody chip kit for diagnosis of various tumors
  • Antibody chip kit for diagnosis of various tumors
  • Antibody chip kit for diagnosis of various tumors

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 diagnosis of various tumors according to the present invention.

[0028] In order to detect whether there are corresponding tumor markers in the sample, prepare slides immobilized with specific antibodies against the following tumor markers: alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), cancer antigen CA125, cancer antigen CA19 -9, cancer antigen CA153, prostate-specific antigen (fPSA and tPSA), neuron-specific enolase (NSE), calcitonin (PCT), iron-binding protein (Ferritin), β2-microglobulin (beta 2 -Microglobin), human chorionic gonadotropin beta (HCGb), pepsinogen (Pepsinogen 1 and 2), thyroglobulin (Thyroglobulin), prolactin and human epididymis secretory protein 4 (HE4).

[0029] 1. The source of the antibody:

[0030] Specific antibodies against the proteins listed in Table 1 were used, and the use, source, and concentration of the antibodies are detailed in Table 1. All detection antibodies were lab...

Embodiment 3

[0042] Example 3: An experiment of quantitative detection of tumor markers using the kit of the present invention.

[0043] 1. Complete drying of the slide chip

[0044] 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.

[0045] 2. Gradient dilution of tumor marker standards

[0046] 2.1 Add 500 μl of sample diluent to the vial of the tumor 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 Std 1.

[0047] 2.2 Mark six clean centrifuge tubes as Std2, Std3 to Std7 respectively, and add 200 μl of sample diluent to each small tube.

[0048] 2.3 Take 100 μl of Std 1 and add it to Std2 and mix gently, then take 100 μl from Std 2 and add it to Std 3, and then dilute to ...

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Abstract

The invention relates to an antibody chip kit for diagnosis of various tumors. The kit includes an antibody chip, a tumor marker standard substance mixture, a biotin-labeled tumor marker detection antibody mixture, and a fluorescein Cy3-labeled streptavidin, wherein the antibody chip includes a substrate, 16 kinds of tumor marker specific antibodies fixed on the surface of the substrate, and two kinds of positive controls, and the tumor marker standard substance mixture is a freeze-dried mixture obtained by mixing 16 kinds of standard tumor marker standard substances together according to a certain amount. The kit can detect 16 clinical commonly used tumor markers, overcomes the defects of complicated operation, single detection index, low sensitivity and the like in the prior art, has the advantages of being cheap, convenient, sensitive, accurate, high in throughput, and less in amount of samples, and can be popularized and large-scaled in common laboratories.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an antibody chip kit for diagnosis of various tumors. Background technique [0002] Since the 21st century, the threat of malignant tumors to human health has become increasingly serious. Its fatality rate is second only to cardiovascular and cerebrovascular diseases, ranking third. The main reason for the high mortality of cancer patients is the inaccurate diagnosis. Accurate diagnosis and timely treatment are the most effective ways to prevent and treat cancer and reduce mortality. [0003] With the development of tumor cells, certain proteins in the body of tumor patients will change, or new abnormal proteins associated with tumors will be produced. These chemical substances that can reflect the presence of tumors are collectively called tumor markers. Since tumor markers either do not exist in normal adult tissues but are only found in tumor tissues, or their content i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57484
Inventor 黄若磐毛应清
Owner RAYBIOTECH INC GUANGZHOU
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