CN109470847A - A kind of antibody chip data are extracted and analysis method - Google Patents
A kind of antibody chip data are extracted and analysis method Download PDFInfo
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- CN109470847A CN109470847A CN201811314427.3A CN201811314427A CN109470847A CN 109470847 A CN109470847 A CN 109470847A CN 201811314427 A CN201811314427 A CN 201811314427A CN 109470847 A CN109470847 A CN 109470847A
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- antibody chip
- photo
- developing apparatus
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- extracted
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- 102000004965 antibodies Human genes 0.000 title claims abstract description 68
- 108090001123 antibodies Proteins 0.000 title claims abstract description 68
- 238000004458 analytical method Methods 0.000 title claims abstract description 27
- 230000011664 signaling Effects 0.000 claims abstract description 24
- 230000000875 corresponding Effects 0.000 claims abstract description 17
- 238000001514 detection method Methods 0.000 claims abstract description 11
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 9
- 238000004020 luminiscence type Methods 0.000 claims abstract description 7
- 238000003908 quality control method Methods 0.000 claims description 7
- 238000007405 data analysis Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 10
- UIIMBOGNXHQVGW-UHFFFAOYSA-M buffer Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 9
- 238000000034 method Methods 0.000 description 5
- 238000004140 cleaning Methods 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000011534 wash buffer Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 239000006210 lotion Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000001131 transforming Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 241000208340 Araliaceae Species 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000038129 antigens Human genes 0.000 description 1
- 108091007172 antigens Proteins 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 235000005035 ginseng Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
Abstract
The present invention relates to a kind of antibody chip reading data and analysis methods, comprising the following steps: a complete antibody chip of incubation is placed in a developing apparatus by step 1, and luminescence reagent is added on the surface of the experimental film of the antibody chip；Step 2 takes pictures to the antibody chip using the developing apparatus, obtains a plurality of photos with different exposure parameters；The photo of step 3, selection one with proper exposure parameter opens the photo using analysis software；Step 4 is selected a plurality of signaling points on the photo using the analysis software, and reads the corresponding OD value of the signaling point using the analysis software；Step 5 carries out analysis detection to the signaling point and the corresponding OD value；Wherein, the developing apparatus is ChemiScope 6000, and the analysis software is HLImage.The advantage is that, by using ChemiScope 6000 and HLImage, to various antibody chip kits carry out reading data analysis, improve the accuracy and repeatability of experimental data.
Description
Technical field
The present invention relates to technical field of molecular biology more particularly to a kind of antibody chip data are extracted and analysis method.
Background technique
Antibody chip have the characteristics that micromation, it is integrated, high-throughput, can be used in detecting a certain specific physiology or disease
The gene expression abundance of reason process GAP-associated protein GAP, is mainly used for the correlative study of signal transduction, protein science, tumour and other diseases.
Antibody chip will be fixed on carrier to high-density from the Multiple Antibodies in conjunction with different antigentic specificities, be made to be measured
Sample washes off the albumen of non-specific binding by elution by chip surface, thus to specific binding in antibody chip
The antigen on surface is detected.
However for existing antibody chip detection kit, since each kit reagent is different, detection method is different, leads
It causes in detection process, experimental data is inaccurate, and can not preferably carry out repeated experiment.
Therefore, a kind of detection method that can be suitable for different antibodies chip inspecting reagent unit is needed, the essence of data is improved
True property and repeatability.
Summary of the invention
The purpose of the present invention is aiming at the shortcomings in the prior art, provide a kind of antibody chip data to extract and analysis side
Method.
To achieve the above object, the technical solution adopted by the present invention is that:
A kind of antibody chip data are extracted and analysis method, comprising the following steps:
The one complete antibody chip of incubation is placed in a developing apparatus by step 1, in the experimental film of the antibody chip
Surface be added luminescence reagent；
Step 2 takes pictures to the antibody chip using the developing apparatus, obtains a plurality of with different exposures ginseng
Several photos；
The photo of step 3, selection one with proper exposure parameter opens the photo using analysis software；
Step 4 is selected a plurality of signaling points on the photo using the analysis software, and uses the analysis software
Read the corresponding OD value of the signaling point；
Step 5 carries out analysis detection to the signaling point and the corresponding OD value；
Wherein, the developing apparatus is ChemiScope 6000, and the analysis software is HLImage.
Preferably, it in the step 2, before being taken pictures using the developing apparatus to the antibody chip, adjusts and claps
According to exposure mode.
Preferably, the exposure mode is automatic exposure or Manual exposure.
Preferably, in the step 1, after the antibody chip is placed in the developing apparatus, the development is adjusted
The focal length of device.
Preferably, the luminescence reagent is ECL luminescent solution.
Preferably, in the step 3, select the method for the photo with proper exposure parameter for according to the antibody core
The development situation of the Quality Control point of on piece selects the Quality Control point not occur the photo of burst point.
The invention adopts the above technical scheme, compared with prior art, has the following technical effect that
A kind of antibody chip data of the invention are extracted and analysis method, by using 6000 He of ChemiScope
HLImage, the experiment that can be carried out to existing various antibody chip kits carry out reading data and analysis, improve experiment
The accuracy and repeatability of data, while keeping reading data and analysis more efficient and convenient.
Detailed description of the invention
Fig. 1 is the photo of antibody chip in the prior art.
Fig. 2 is the photo of antibody chip of the invention.
Fig. 3 is the signaling point schematic diagram of the antibody chip of one embodiment of the present of invention.
Fig. 4 is the signaling point schematic diagram of the antibody chip of one embodiment of the present of invention.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, those of ordinary skill in the art without creative labor it is obtained it is all its
His embodiment, shall fall within the protection scope of the present invention.
It should be noted that in the absence of conflict, the feature in embodiment and embodiment in the present invention can phase
Mutually combination.
The present invention is further explained in the light of specific embodiments, but not as the limitation of the invention.
Embodiment 1
Of the invention a kind of antibody chip reading data and analysis method, comprising the following steps:
The one complete antibody chip of incubation is placed in a developing apparatus by step 1, in the table of the experimental film of antibody chip
Luminescence reagent is added in face；
Step 2 takes pictures to antibody chip using developing apparatus, obtains a plurality of photographs with different exposure parameters
Piece；
The photo of step 3, selection one with proper exposure parameter opens photo using analysis software；
Step 4 is selected a plurality of signaling points on photo using analysis software, and reads signaling point pair using analysis software
The OD value answered；
Step 5 carries out analysis detection to signaling point and corresponding OD value；
Wherein, developing apparatus is ChemiScope 6000, and analysis software is HLImage.
Further, in step 1, after antibody chip being placed in the exposure device of developing apparatus, development dress is adjusted
The focal length set.
Further, in step 1, the luminescence reagent of addition is ECL luminescent solution.
Further, in step 2, the exposure taken pictures before developing apparatus takes pictures to antibody chip, to developing apparatus
Mode is adjusted, to obtain the photo of different exposure parameters.
Further, in step 2, exposure mode is automatic exposure or Manual exposure.
Further, in step 3, select the method for the photo with proper exposure parameter for according to the matter of antibody chip
The development situation for controlling point, selects Quality Control point not occur the photo of burst point.
In the present invention, developing apparatus selection ChemiScope 6000 (Shanghai Qin Xiang scientific instrument Co., Ltd), it is sensitive
Degree is high, image taking speed is fast, the range of linearity is wide；Result and automatic whole can be analyzed with automatic identification by analyzing software selection HLImage
Reason analysis data.
Same antibody chip is subjected to comparison of taking pictures using the prior art and method of the invention, as shown in Fig. 1~2,
Positobe focus overexposure in Fig. 1 of prior art shooting, leads to not preferably read corresponding data, and shooting of the invention
Fig. 2 then without overexposure phenomenon, can preferably read corresponding data.
Embodiment 2
The present embodiment is a specific embodiment of the invention.
The one complete antibody chip (the model RnD-Ary005B of antibody chip) of incubation is placed in by step 1
In ChemiScope 6000, ECL luminescent solution is added on the surface of the experimental film of antibody chip；
Step 2 takes pictures to the antibody chip using ChemiScope 6000, and the exposure mode selected exposes to be automatic
Light and the high voxel model of selection, obtain multiple photos with different exposure parameters；
One step 3, selection photo with proper exposure parameter, i.e. Quality Control point on selection antibody chip do not occur quick-fried
The photo of point opens photo using HLImage；
Step 4 is selected a plurality of signaling points automatically on photo using HLImage, and is read automatically often using HLImage
The corresponding OD value of a signaling point；
Step 5 carries out analysis detection to each signaling point and corresponding OD value.
Specifically, the incubation processing of the antibody chip of step 1 is as follows:
Sample size determines: cells and supernatant volume 700ul；
Closed protein chip: every film needs to do Seal treatment with the Array Buffer 4 of 2ml, and 1 is incubated on shaking table
Hour；
The processing of sample: need to be diluted 0.5ml sample with 1ml Array Buffer 4 to 1.5ml, if volume is insufficient
1.5ml then needs to be adjusted with Array Buffer 5 to 1.5ml；
The detection antibody of 15ul is added in each sample, is incubated for 1 hour, incubation temperature is 4 DEG C；
Array Buffer 4 is sucked, the sample that preparation is added is respectively corresponded；
Shaker overnight is incubated for, and temperature is at 2-8 DEG C；
It is cleaned experimental film 3 times of antibody chip with the 1* washing buffer of 20ml, cleans 10min every time；
Streptavidin-the HRP of 2ml is added, is incubated for 30min, shaking table vibrates under room temperature；
It is cleaned experimental film 3 times of antibody chip with the 1* washing buffer of 20ml again, cleans 10min every time；
Most of washing lotion on experimental film is sucked, the ECL luminescent solution of 1ml in incubation, when incubation reduces bubble production to the greatest extent
It is raw, it is incubated for about 1min；
Suck ECL luminescent solution.
Specifically, the detailed process of HLImage analysis are as follows:
The format of photo is converted into bmp format, then imports the photo after format transformation；
Corresponding antibody chip type (i.e. RnD-Ary005B) is selected in the analysis module of HLImage, is then carried out certainly
Dynamic identification；
Staff according to the actual situation, is adaptively adjusted identification aperture, final image is as shown in Figure 3；
HLImage reads the OD value of each signaling point, and all data are exported, and export is excel format
File；
Signaling point and corresponding OD value are analyzed.
Embodiment 3
The present embodiment is a specific embodiment of the invention.
The one complete antibody chip (the model RnD-ARY026 of antibody chip) of incubation is placed in by step 1
In ChemiScope 6000, ECL luminescent solution is added on the surface of the experimental film of antibody chip；
Step 2 takes pictures to the antibody chip using ChemiScope 6000, and the exposure mode selected is exposure manually
Light and the high voxel model of selection, obtain multiple photos with different exposure parameters；
One step 3, selection photo with proper exposure parameter, i.e. Quality Control point on selection antibody chip do not occur quick-fried
The photo of point opens photo using HLImage；
Step 4 is selected a plurality of signaling points automatically on photo using HLImage, and is read automatically often using HLImage
The corresponding OD value of a signaling point；
Step 5 carries out analysis detection to each signaling point and corresponding OD value.
Specifically, the incubation processing of the antibody chip of step 1 is as follows:
Sample size determines: every membrane sample 500ul；
Closed protein chip: every film needs to do Seal treatment with the Array Buffer 6 of 2ml, and 1 is incubated on shaking table
Hour；
The processing of sample: need to be diluted 0.5ml sample with 1ml Array Buffer 6 to 1.5ml；
Array Buffer 6 is sucked, the sample that preparation is added is respectively corresponded；
Shaker overnight is incubated for, and temperature is at 2-8 DEG C；
It is cleaned experimental film 3 times of antibody chip with the 1* washing buffer of 20ml, cleans 10min every time；
For every film, 30ul is diluted with Array Buffer 4/6 and detects antibody to 1.5ml, and every film is added and incubates
It educates, shaking table is incubated for 1h；
Streptavidin-the HRP (Array Buffer 2/3 is according to dilution) of 2ml is added, is incubated for 30min, room
Shaking table vibrates under the conditions of temperature；
It is cleaned experimental film 3 times of antibody chip with the 1* washing buffer of 20ml again, cleans 10min every time；
Most of washing lotion on experimental film is sucked, the ECL luminescent solution of 1ml in incubation, when incubation reduces bubble production to the greatest extent
It is raw, it is incubated for about 1min；
Suck ECL luminescent solution.
Specifically, the detailed process of HLImage analysis are as follows:
The format of photo is converted into bmp format, then imports the photo after format transformation；
Corresponding antibody chip type (i.e. RnD-ARY026) is selected in the analysis module of HLImage, is then carried out certainly
Dynamic identification；
Staff according to the actual situation, is adaptively adjusted identification aperture, final image is as shown in Figure 4；
HLImage reads the OD value of each signaling point, and all data are exported, and export is excel format
File；
Signaling point and corresponding OD value are analyzed.
The foregoing is merely preferred embodiments of the present invention, are not intended to limit embodiments of the present invention and protection model
It encloses, to those skilled in the art, should can appreciate that all with made by description of the invention and diagramatic content
Equivalent replacement and obviously change obtained scheme, should all be included within the scope of the present invention.
Claims (6)
1. a kind of antibody chip data are extracted and analysis method, which comprises the following steps:
The one complete antibody chip of incubation is placed in a developing apparatus by step 1, in the table of the experimental film of the antibody chip
Luminescence reagent is added in face；
Step 2 takes pictures to the antibody chip using the developing apparatus, obtains a plurality of with different exposure parameters
Photo；
The photo of step 3, selection one with proper exposure parameter opens the photo using analysis software；
Step 4 is selected a plurality of signaling points on the photo using the analysis software, and is read using the analysis software
The corresponding OD value of the signaling point；
Step 5 carries out analysis detection to the signaling point and the corresponding OD value；
Wherein, the developing apparatus is ChemiScope 6000, and the analysis software is HLImage.
2. antibody chip data according to claim 1 are extracted and analysis method, which is characterized in that in the step 2,
Before taking pictures using the developing apparatus to the antibody chip, the exposure mode taken pictures is adjusted.
3. antibody chip data according to claim 2 are extracted and analysis method, which is characterized in that the exposure mode is
Automatic exposure or Manual exposure.
4. antibody chip data according to claim 1 are extracted and analysis method, which is characterized in that in the step 1,
After the antibody chip is placed in the developing apparatus, the focal length of the developing apparatus is adjusted.
5. antibody chip data according to claim 1 are extracted and analysis method, which is characterized in that the luminescence reagent is
ECL luminescent solution.
6. antibody chip data according to claim 1 are extracted and analysis method, which is characterized in that in the step 3,
The method of the photo with proper exposure parameter is selected to select institute according to the development situation of the Quality Control point on the antibody chip
It states Quality Control point and does not occur the photo of burst point.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811314427.3A CN109470847A (en) | 2018-11-06 | 2018-11-06 | A kind of antibody chip data are extracted and analysis method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811314427.3A CN109470847A (en) | 2018-11-06 | 2018-11-06 | A kind of antibody chip data are extracted and analysis method |
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|---|---|
| CN109470847A true CN109470847A (en) | 2019-03-15 |
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| CN201811314427.3A Pending CN109470847A (en) | 2018-11-06 | 2018-11-06 | A kind of antibody chip data are extracted and analysis method |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100060856A1 (en) * | 2008-09-11 | 2010-03-11 | Photo Me Holding France | Installation for exposing a cinematographic film from digital images |
| CN103217541A (en) * | 2013-04-23 | 2013-07-24 | 上海裕隆生物科技有限公司 | Full-automatic pipe-type chemiluminescent analyzer |
| CN105334325A (en) * | 2015-11-12 | 2016-02-17 | 国家纳米科学中心 | Microfluidic immune chip analysis method based on biotin and streptavidine system |
| CN108387750A (en) * | 2018-02-08 | 2018-08-10 | 江苏三联生物工程有限公司 | A kind of chip and preparation method thereof of joint-detection F-T3, F-T4 and TSH |
| CN108548780A (en) * | 2018-03-30 | 2018-09-18 | 中国人民解放军第二军医大学 | The method of transcription factor chip agent box and high flux screening target gene transcription factor |
-
2018
- 2018-11-06 CN CN201811314427.3A patent/CN109470847A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100060856A1 (en) * | 2008-09-11 | 2010-03-11 | Photo Me Holding France | Installation for exposing a cinematographic film from digital images |
| CN103217541A (en) * | 2013-04-23 | 2013-07-24 | 上海裕隆生物科技有限公司 | Full-automatic pipe-type chemiluminescent analyzer |
| CN105334325A (en) * | 2015-11-12 | 2016-02-17 | 国家纳米科学中心 | Microfluidic immune chip analysis method based on biotin and streptavidine system |
| CN108387750A (en) * | 2018-02-08 | 2018-08-10 | 江苏三联生物工程有限公司 | A kind of chip and preparation method thereof of joint-detection F-T3, F-T4 and TSH |
| CN108548780A (en) * | 2018-03-30 | 2018-09-18 | 中国人民解放军第二军医大学 | The method of transcription factor chip agent box and high flux screening target gene transcription factor |
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