CN106248959B - A kind of immunodiafiltration method and immunodiafiltration device for detecting serum fucosin - Google Patents
A kind of immunodiafiltration method and immunodiafiltration device for detecting serum fucosin Download PDFInfo
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Abstract
本发明涉及一种检测血清岩藻糖蛋白的免疫渗滤方法及免疫渗滤装置，所述方法包括如下步骤：(1)将甲胎蛋白抗原和小扁豆素复合于微孔滤膜上；(2)滴加血清样品到微孔滤膜上，使样品中岩藻糖基化甲胎蛋白与微孔滤膜上的小扁豆素结合；(3)向微孔滤膜上滴加酶标记的甲胎蛋白多克隆抗体，并加入化学发光底物，得到检测结果；本发明还提供了用于检测血清岩藻糖蛋白的免疫渗滤装置。本发明利用微孔滤膜，在较短时间内实现了血清样本中多种抗原或抗体的一次性同时检测，其检测过程只需3‑5min即可完成，具有检测时间短、灵成本低、稳定性好等优点。
The present invention relates to an immunodiafiltration method and an immunodiafiltration device for detecting serum fucose protein. The method comprises the following steps: (1) compounding alpha-fetoprotein antigen and lentil on a microporous filter membrane; ( 2) drop the serum sample on the microporous membrane, so that the fucosylated alpha-fetoprotein in the sample is combined with the lentil on the microporous membrane; The alpha-fetoprotein polyclonal antibody is added, and the chemiluminescence substrate is added to obtain the detection result; the invention also provides an immunodiafiltration device for detecting serum fucosin. The invention utilizes the microporous filter membrane to realize one-time simultaneous detection of multiple antigens or antibodies in serum samples in a relatively short period of time. Good stability and so on.
Description
Technical field
The present invention relates to protein detection technology field more particularly to a kind of immunity percolation sides for detecting serum fucose albumen
Method and immunity percolation device.
Background technique
Alpha-fetoprotein that the benign hepatopathy such as primary carcinoma of liver and hepatitis, cirrhosis generates (alpha fetoprotein,
AFP) there are great differences on its sugar chain structure, i.e., compared with benign hepatopathy, the AFP that liver cancer generates, fucose index is wanted
It is much higher.Fucose has the characteristic in conjunction with lens element.AFP is according to its (fucosido) to the affine of LcA
Power difference can be divided into AFP-L1, AFP-L2 and AFP-L3.Wherein AFP-L1 mostlys come from benign hepatopathy, and AFP-L2 mainly comes
Derived from pregnant woman, and AFP-L3 is the fucose glycoforms of alpha-fetoprotein, is mainly derived from hepatocellular carcinoma
(HepatoCellular Carcinoma, HCC).FDA official approval is using AFP-L3 as the mark of primary carcinoma of liver within 2005
One of object.AFP-L3 the early diagnosis of liver cancer, antidiastole, curative effect evaluation and in terms of all have higher spy
Anisotropic and sensibility.
Fucose is methylation hexose, is present in a variety of glycoprotein candy chains such as tissue and serum, referred to as protein binding
Fucose (protein-bound fucose, P-bf).There are fucosyl residues, such heteroplasmons on AFP carbohydrate chain
Referred to as fucosylation AFP (FucAFP) has important theory significance and clinical application significance, answers in diagnosing cancer of liver and prognosis
It can be used as an important index in.
The separation method of traditional serum fucose albumen, including affine crossed immunoelectrophoresis technology, affine blotting, parent
With chromatography, " two-site sandwich " enzyme linked immunosorbent assay, LiBASys analyzer,I30 detection system technology
And the glycosyl of hot scape biology captures centrifugal column pretreatment technology.Wherein the affine immuno-electrophoresis of phytolectin andI30 detection system technical requirements are high, cumbersome, expensive reagents, limit its popularization and application.And glycosyl is caught
Centrifugal column is obtained, since sample process and detection separately carry out, increases the triviality of operation.
Summary of the invention
The problem of for current fucose protein separating method, the purpose of the present invention is to provide a kind of detection blood
The immunity percolation method and immunity percolation device of clear fucose albumen, the method and immunity percolation device are particularly suitable for detecting
Fucosylation alpha-fetoprotein in biological sample, and there is versatility to various fucosylation albumen.
For this purpose, the invention adopts the following technical scheme:
In a first aspect, the present invention provides a kind of immunity percolation methods for detecting serum fucose albumen comprising as follows
Step:
(1) alpha-fetoprotein antigen and lens element are compound on miillpore filter；
(2) it is added dropwise on blood serum sample to miillpore filter, makes on the fucosylation alpha-fetoprotein and miillpore filter in sample
Lens element combine；
(3) the alpha-fetoprotein polyclonal antibody of enzyme label is added dropwise on miillpore filter, and chromogenic substrate is added, is detected
As a result.
The present invention is to realize the disposable of a variety of antigens in serum sample or antibody in a short time using miillpore filter
It detects simultaneously, detection process only needs 3-5min can be completed.Compared to protein chip is used, immunity percolation provided by the invention is examined
Survey method has many advantages, such as that detection time is short, at low cost, stability is good.
Immunity percolation method of the invention is will to capture proteopexy on nitrocellulose filter, negatively charged antibody with
Being combined together for hydrophobic effect and high-affinity occurs for nitrocellulose filter；Also, occur by using HRP and chromogenic substrate
Reaction generates blue brown stable product, the product can long-time stable exist, will not gradually die down with the extension of time or
It disappears, observation result will not generate error because of the different of period, and as a result stability is good；Further more, immunity percolation of the invention
Method does not need any instrument and carries out result scanning analysis, and judging result can be visually observed within 5min；Secondly, of the invention
Immunity percolation method need to only be completed in room temperature, not need incubator.
Immunity percolation detection method provided by the invention, suitable for detecting the fucosylation first tire egg biological sample
It is white, and there is versatility to various fucosylation albumen.
Fucosylation alpha-fetoprotein in the present invention refers to that there are fucosyl residues on alpha-fetoprotein carbohydrate chain
Heteroplasmon.
According to the present invention, step (1) the alpha-fetoprotein antigen is prokaryotic expression human a-fetoprotein antigen, also be may be selected true
Nuclear expression alpha-fetoprotein antigen, does not do particular determination herein.
According to the present invention, the alpha-fetoprotein polyclonal antibody of step (3) the enzyme label is rabbit source antibody.
According to the present invention, step (3) enzyme is horseradish peroxidase (HRP), can also be alkaline phosphatase (ALP),
It is for realizing chromogenic enzyme substrate.
According to the present invention, the immunity percolation method of the detection serum fucose albumen may include steps of:
(1) alpha-fetoprotein antigen and lens element are compound on miillpore filter；
(2) be added dropwise on the blood serum sample to miillpore filter of 10-15 μ L, make fucosylation alpha-fetoprotein in sample with it is micro-
Lens element on the filter membrane of hole combines；Then it is washed with confining liquid, removes non-specific binding thing；
(3) the alpha-fetoprotein polyclonal antibody of enzyme label is added dropwise on miillpore filter, is washed after being incubated for 3-5s with confining liquid,
Non-specific binding thing is removed, chromogenic substrate developing solution is added；
When positive serum, the detection zone of alpha-fetoprotein antigen and fucosylation alpha-fetoprotein is displayed in blue, negative blood
When clear, alpha-fetoprotein antigen detection zone is displayed in blue, and the detection zone of fucosylation alpha-fetoprotein does not develop the color.Negative control
Either positive serum and negative serum do not develop the color.
Illustratively, the immunity percolation method can specifically include following steps:
Sample detection:
It will be added dropwise on the detection zone containing miillpore filter after the dilution of test serum sample, after incubation, wash inspection with PBST
Area is surveyed, non-specific binding object is removed；The AFP antibody with the diluted HRP label of PBS is added, after incubation, is washed, is removed with PBST
Non-specific binding object；HRP substrate developing solution, visual results are added.
According to the present invention, the incubation refers to incubation at room temperature 3-5s, such as 3s, 3.5s, 4s, 4.5s or 5s.
It according to the present invention, can be in following immunity percolation in the immunity percolation method of the detection serum fucose albumen
It is carried out in device, but is not limited only to this.
A kind of immunity percolation device detecting serum fucose albumen, containing miillpore filter, in miillpore filter matrix
Be provided at least one detection zone, be equipped in the detection zone positive control detection subprovince, serum fucose Protein Detection subprovince and
Negative control area, wherein positive control detection subprovince is fixed with alpha-fetoprotein, and serum fucose Protein Detection subprovince is fixed with small
Hyacinth bean element, negative control area is fixed with bovine serum albumin(BSA)；The material concentration on detection spot in same detection subprovince is identical.Inspection
It surveys subprovince and check plot includes at least 1 detection spot respectively, such as can be 2,3,4,5 or 6 etc..
Heretofore described immunity percolation device by good toughness plastic products, papery water-absorbent material and nitrocellulose filter
Composition, is able to bear shock and collision, it is not easy to broken.
In the present invention, there is 1 detection zone in the miillpore filter matrix, the detection subprovince has 1 detection spot, institute
Stating positive control area and negative control area respectively has 1 control spot；The alpha-fetoprotein is prokaryotic expression human a-fetoprotein.
Immunity percolation device of the invention further includes alpha-fetoprotein polyclonal antibody and the colour developing of horseradish peroxidase label
Substrate solution；The alpha-fetoprotein polyclonal antibody of the horseradish peroxidase label is rabbit source antibody；The immunity percolation device is also
Including for washing and diluted PBST and PBS confining liquid.
Present invention preferably employs the immunity percolation devices with miillpore filter, compared with protein chip, when having detection
Between it is short, at low cost, stability is good the advantages that, process control can be will test in 3-5min.
Second aspect, it is described immune the present invention also provides a kind of immunity percolation device for detecting serum fucose albumen
Include at least one detection zone in the miillpore filter matrix of percolating device, is equipped with positive control in the detection zone and detects subprovince, blood
Clear fucose Protein Detection subprovince and negative control area, wherein positive control detection subprovince is fixed with alpha-fetoprotein, serum rock algae
Glycoprotein detection subprovince is fixed with lens element, and negative control area is fixed with bovine serum albumin(BSA)；Inspection in same detection subprovince
The material concentration surveyed on spot is identical.
According to the present invention, the detection subprovince includes at least 1 detection spot, such as can be 2,3,4,5 or 6
It is a etc..
According to the present invention, there is 1 detection zone in the miillpore filter matrix, the detection subprovince has 1 detection spot,
The positive control area and negative control area respectively have 1 control spot.
According to the present invention, the alpha-fetoprotein is prokaryotic expression human a-fetoprotein.
According to the present invention, the immunity percolation device further includes the alpha-fetoprotein polyclonal antibody of horseradish peroxidase label
And assay chromogenic substrate solution.
According to the present invention, the alpha-fetoprotein polyclonal antibody of the horseradish peroxidase label is rabbit source antibody.
According to the present invention, the immunity percolation device further includes for washing and diluted PBST and PBS confining liquid.
The present invention also provides immunity percolation devices in non-diagnostic and/or non-treatment purpose serum fucose Protein Detection
Purposes.
Compared with prior art, the present invention is at least had the advantages that
(1) present invention is by utilizing miillpore filter, realizes a variety of antigens in serum sample or antibody in a short time
It disposably detects simultaneously, immunity percolation detection method provided by the invention can accurately detect serum fucose albumen, have
The advantages that detection time is short, stability is good, at low cost；
(2) raw material and instrument and equipment cost that detection method provided by the invention and immunity percolation device need are all greatly
It reduces, the device of immunity percolation is papery and plastic products, and cost is very low, about 1 yuan, does not need any instrument and equipment
Preparation, incubation, the result scanning for carrying out experiment flow, greatly reduce testing cost and expense；
(3) immunity percolation method of the invention is compared with other methods in time, and ELISA detection at least needs
3h；Protein chip detection needs 1.5h；And the present invention only needs 3-5min, can greatly shorten in detection time.
Detailed description of the invention
Fig. 1 is immunity percolation schematic device of the invention, wherein 1- lid, 2- miillpore filter, and 3- papery water-absorbent material,
4- plastic bottom.
Fig. 2 is the result schematic diagram of immunity percolation method of the invention, wherein 1-3 is positive findings, the sun above NC film
Property control test spot blue brown positive signal is presented, blue brown positive signal is presented in intermediate lens element detection spot, lower section
BSA control spot does not develop the color；4-5 is negative findings, and blue brown positive signal is presented in the positive control detection spot above NC film, intermediate
Lens element detection spot and lower section BSA control spot do not develop the color.
Fig. 3 is the schematic diagram for preparing immunity percolation nitrocellulose filter, and the fixed AFP in positive control area, intermediate detection area consolidates
Determine lens element, the fixed BSA in negative control area.
The present invention is described in more detail below.But following examples is only simple example of the invention, not generation
Table or limitation the scope of the present invention, protection scope of the present invention are subject to claims.
Specific embodiment
To further illustrate the technical scheme of the present invention below with reference to the accompanying drawings and specific embodiments.
As shown in Figure 1, it is the structural schematic diagram of immunity percolation device in the present invention, which has micropore
Filter membrane is set gradually from top to bottom as lid 1, miillpore filter 2, papery water-absorbent material 3 and plastic bottom 4.In assembling, in advance will
Miillpore filter is cut into the circle of diameter about 1.5cm, and papery absorbing material 3 is placed in plastic bottom 4, on papery water-absorbent material
The miillpore filter 2 that cuts in advance is placed in face center, then closes the lid 1.
Used miillpore filter can be nitrocellulose filter in the present invention, and the present invention uses GE Healthcare
RPN303B；Papery water-absorbent material can be blotting paper；Lid and bottom are all made of plastic products.
As shown in Fig. 2, it illustrates the result schematic diagram using immunity percolation method of the invention, (Fig. 2 when positive serum
In 1-3), the detection zone of positive control and fucosylation alpha-fetoprotein is displayed in blue；(figure when if it is negative serum
4-6 in 2), the detection zone of fucosylation alpha-fetoprotein does not develop the color then, and positive control is displayed in blue；It is no matter positive and negative
Property serum, negative control do not develop the color.
Fig. 3 is the schematic diagram for preparing immunity percolation nitrocellulose filter, and the fixed AFP in positive control area, intermediate detection area consolidates
Determine lens element, the fixed BSA in negative control area.
In order to better illustrate the present invention, it is easy to understand technical solution of the present invention, of the invention is typical but non-limiting
Embodiment is as follows:
The preparation and process for using of 1 immunity percolation device of embodiment
Test agents useful for same and instrument: prokaryotic expression AFP (abcam company of the U.S.)；Lens element (Sigma company)；Nitre
The rabbit source antibody (abcam company of the U.S.) of acid cellulose film (GE Healthcare), horseradish peroxidase-labeled；HRP colour developing
Substrate solution (Millipore company of the U.S.).
PBS formula: sodium chloride (NaCl) 8g, potassium chloride (KCl) 0.2g, disodium hydrogen phosphate (Na2HPO4) 1.44g, di(2-ethylhexyl)phosphate
Hydrogen potassium (KH2PO4) 0.24g, it adjusts pH value to 7.4, is settled to 1L.
PBST formula: PBS, 1L+Tween-20,1mL.
Nitrocellulose filter (GE Healthcare), each immunity percolation device include 1 detection zone, each detection zone
Detectable portion serum, one-time detection AFPL3 marker.
In each detection zone, by source of mouse prokaryotic expression people AFP (abcam company of the U.S.), lens element Sigma company)
Each 0.5 μ L is successively put on nitrocellulose filter, point sample concentration AFP 0.5mg/mL, lens element 4mg/mL；10% cow's serum
Albumin (BSA) is used as negative control, and 0.5 μ L point shines spot in pairs.
Immunity percolation operating process:
With AFP L3 tumour in the immunity percolation detection healthy control group and liver cancer experimental group dynamic serum specimen prepared
Marker.
Under room temperature, it is added dropwise on the diaphragm of immunity percolation device with 10 μ L of serum sample, utilizes lens element and rock
The characteristic that algae sugar combines, makes lens element form lens element antigenic compound in conjunction with the fucose in serum；After 3 seconds,
Sera liquid permeates the water-absorbent material water suction below diaphragm envelope piece.
10 μ L PBST are added dropwise to wash, removal non-specific binding, 3 times repeatedly.Add the diluted horseradish peroxidase of PBS
Enzyme marks rabbit source primary antibody, the AFP antigen binding in rabbit source antibody and serum, lens element-(AFP) fucose-rabbit source horseradish mistake
Oxide enzymic-labelled antibody compound；Rabbit source antibody is simultaneously in conjunction with the positive control AFP on fixed film, AFP- rabbit source horseradish mistake
Oxide enzymic-labelled antibody compound.After 3 seconds, sera liquid permeates the water-absorbent material water suction below diaphragm envelope piece.
10 μ L PBST washing is added dropwise, removes non-specific binding, 3 times repeatedly, HRP chromogenic substrate is added, is protected from light 1 minute, drip
10 μ L PBST are added to wash.Positive serum detects spot and blue spot is presented.
Embodiment 2
With AFP L3 tumour in the immunity percolation detection healthy control group and liver cancer experimental group dynamic serum specimen prepared
Marker.
Under room temperature, it after diluting 4 times with 2.5 μ L of serum sample, is added dropwise on the diaphragm of immunity percolation device, utilization is small
The characteristic that hyacinth bean element and fucose combine, makes lens element form lens element antigen in conjunction with the fucose in serum compound
Object；After 5 seconds, sera liquid permeates the water-absorbent material water suction below diaphragm envelope piece.
10 μ L PBST are added dropwise to wash, removal non-specific binding, 5 times repeatedly.Add the diluted horseradish peroxidase of PBS
Enzyme marks rabbit source primary antibody, the AFP antigen binding in rabbit source antibody and serum, lens element-(AFP) fucose-rabbit source horseradish mistake
Oxide enzymic-labelled antibody compound；Rabbit source antibody is simultaneously in conjunction with the positive control AFP on fixed film, AFP- rabbit source horseradish mistake
Oxide enzymic-labelled antibody compound.After 5 seconds, sera liquid permeates the water-absorbent material water suction below diaphragm envelope piece.
10 μ L PBST washing is added dropwise, removes non-specific binding, 5 times repeatedly, HRP chromogenic substrate is added, is protected from light 1 minute, drip
10 μ L PBST are added to wash.Positive serum detects spot and blue spot is presented.
It in summary it can be seen, the present invention utilizes miillpore filter, realizes a variety of antigens in serum sample within a short period of time
Or the disposable of antibody detects simultaneously, detection process only needs 3-5min can be completed, have detection time it is short, spirit it is at low cost,
The advantages that stability is good.
The Applicant declares that the present invention is explained by the above embodiments detailed construction feature of the invention, but the present invention is simultaneously
It is not limited to above-mentioned detailed construction feature, that is, does not mean that the present invention must rely on above-mentioned detailed construction feature and could implement.Institute
Belong to those skilled in the art it will be clearly understood that any improvement in the present invention, to the equivalence replacement of component selected by the present invention
And increase, selection of concrete mode of accessory etc., all of which fall within the scope of protection and disclosure of the present invention.
The preferred embodiment of the present invention has been described above in detail, still, during present invention is not limited to the embodiments described above
Detail within the scope of the technical concept of the present invention can be with various simple variants of the technical solution of the present invention are made, this
A little simple variants all belong to the scope of protection of the present invention.
It is further to note that specific technical features described in the above specific embodiments, in not lance
In the case where shield, can be combined in any appropriate way, in order to avoid unnecessary repetition, the present invention to it is various can
No further explanation will be given for the combination of energy.
In addition, various embodiments of the present invention can be combined randomly, as long as it is without prejudice to originally
The thought of invention, it should also be regarded as the disclosure of the present invention.
Claims (5)
1. a kind of immunity percolation device for detecting serum fucose albumen, which is characterized in that the micropore of the immunity percolation device
Include at least one detection zone in filter membrane matrix, positive control detection subprovince, the inspection of serum fucose albumen are equipped in the detection zone
Subprovince and negative control area are surveyed, wherein positive control detection subprovince is fixed with alpha-fetoprotein, serum fucose Protein Detection subprovince
It is fixed with lens element, negative control area is fixed with bovine serum albumin(BSA)；The substance on detection spot in same detection subprovince is dense
It spends identical；
Wherein, the immunity percolation device further includes that the prokaryotic expression human a-fetoprotein rabbit source of horseradish peroxidase label is polyclonal
Antibody and assay chromogenic substrate solution.
2. immunity percolation device as described in claim 1, which is characterized in that the detection subprovince includes at least 1 detection spot.
3. immunity percolation device as described in claim 1, which is characterized in that have 1 detection in the miillpore filter matrix
Area, the detection subprovince have 1 detection spot, and the positive control area and negative control area respectively have 1 control spot.
4. immunity percolation device as described in claim 1, which is characterized in that the immunity percolation device further includes for washing
With diluted PBST and PBS confining liquid.
5. immunity percolation device as described in claim 1 is examined in non-diagnostic and/or non-treatment purpose serum fucose albumen
Purposes in survey.
Priority Applications (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610579801.7A CN106248959B (en) | 2016-07-21 | 2016-07-21 | A kind of immunodiafiltration method and immunodiafiltration device for detecting serum fucosin |
| AU2016101733A AU2016101733A4 (en) | 2016-07-21 | 2016-09-29 | An immune filtration method and device for detecting serum fucosylated glycoprotein |
| RU2019104468U RU198108U1 (en) | 2016-07-21 | 2017-06-29 | Immunofiltration device for the analysis of fucosylated glycoprotein in blood serum |
| PCT/CN2017/090864 WO2018014709A1 (en) | 2016-07-21 | 2017-06-29 | Immunofiltration method and immunofiltration device for serum fucose protein assay |
| JP2019600070U JP3222251U (en) | 2016-07-21 | 2017-06-29 | Immunodiafiltration apparatus for detecting serum fucose protein |
| DE212017000022.8U DE212017000022U1 (en) | 2016-07-21 | 2017-06-29 | An immunoinfiltration device for detecting fucosylated glycoprotein in serum |
| PH12019500151A PH12019500151A1 (en) | 2016-07-21 | 2019-01-21 | Immunofiltration method and immunofiltration device for serum fucose protein assay |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610579801.7A CN106248959B (en) | 2016-07-21 | 2016-07-21 | A kind of immunodiafiltration method and immunodiafiltration device for detecting serum fucosin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106248959A CN106248959A (en) | 2016-12-21 |
| CN106248959B true CN106248959B (en) | 2019-01-25 |
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| CN201610579801.7A Expired - Fee Related CN106248959B (en) | 2016-07-21 | 2016-07-21 | A kind of immunodiafiltration method and immunodiafiltration device for detecting serum fucosin |
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| JP (1) | JP3222251U (en) |
| CN (1) | CN106248959B (en) |
| AU (1) | AU2016101733A4 (en) |
| DE (1) | DE212017000022U1 (en) |
| PH (1) | PH12019500151A1 (en) |
| RU (1) | RU198108U1 (en) |
| WO (1) | WO2018014709A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106248959B (en) * | 2016-07-21 | 2019-01-25 | 首都医科大学附属北京佑安医院 | A kind of immunodiafiltration method and immunodiafiltration device for detecting serum fucosin |
| CN119574891A (en) * | 2025-02-06 | 2025-03-07 | 杭州浙大迪迅生物基因工程有限公司 | A fluorescent percolation kit for detecting allergen-specific IgE antibodies |
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| EP2950098A1 (en) * | 2009-07-14 | 2015-12-02 | National Institute of Advanced Industrial Science and Technology | Glycan markers as measure of disease state of hepatic diseases |
| US20110223607A1 (en) * | 2010-03-12 | 2011-09-15 | Quest Diagnostics Investments Incorporated | Circulating microrna as a marker for hepatocellular carcinoma |
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2016
- 2016-07-21 CN CN201610579801.7A patent/CN106248959B/en not_active Expired - Fee Related
- 2016-09-29 AU AU2016101733A patent/AU2016101733A4/en not_active Ceased
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2017
- 2017-06-29 JP JP2019600070U patent/JP3222251U/en not_active Expired - Fee Related
- 2017-06-29 DE DE212017000022.8U patent/DE212017000022U1/en not_active Expired - Lifetime
- 2017-06-29 RU RU2019104468U patent/RU198108U1/en active
- 2017-06-29 WO PCT/CN2017/090864 patent/WO2018014709A1/en not_active Ceased
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2019
- 2019-01-21 PH PH12019500151A patent/PH12019500151A1/en unknown
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Also Published As
| Publication number | Publication date |
|---|---|
| RU198108U1 (en) | 2020-06-18 |
| WO2018014709A1 (en) | 2018-01-25 |
| AU2016101733A4 (en) | 2016-10-27 |
| JP3222251U (en) | 2019-07-25 |
| CN106248959A (en) | 2016-12-21 |
| DE212017000022U1 (en) | 2018-02-12 |
| PH12019500151A1 (en) | 2019-10-14 |
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