CN1156702C - Protein chip based on labeling streptavidin-biotin technology - Google Patents
Protein chip based on labeling streptavidin-biotin technology Download PDFInfo
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- CN1156702C CN1156702C CNB011133236A CN01113323A CN1156702C CN 1156702 C CN1156702 C CN 1156702C CN B011133236 A CNB011133236 A CN B011133236A CN 01113323 A CN01113323 A CN 01113323A CN 1156702 C CN1156702 C CN 1156702C
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- 229960002685 biotin Drugs 0.000 title claims abstract description 45
- 239000011616 biotin Substances 0.000 title claims abstract description 45
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 40
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 40
- 238000002372 labelling Methods 0.000 title claims abstract description 26
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims abstract description 40
- 102000004190 Enzymes Human genes 0.000 claims abstract description 36
- 108090000790 Enzymes Proteins 0.000 claims abstract description 36
- 239000000427 antigen Substances 0.000 claims abstract description 33
- 102000036639 antigens Human genes 0.000 claims abstract description 31
- 108091007433 antigens Proteins 0.000 claims abstract description 31
- 238000012360 testing method Methods 0.000 claims abstract description 23
- 235000020958 biotin Nutrition 0.000 claims abstract description 20
- 108010090804 Streptavidin Proteins 0.000 claims abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 16
- 239000000126 substance Substances 0.000 claims abstract description 12
- 238000005406 washing Methods 0.000 claims abstract description 10
- 238000009396 hybridization Methods 0.000 claims abstract description 5
- 239000000020 Nitrocellulose Substances 0.000 claims description 19
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- 239000012528 membrane Substances 0.000 claims description 19
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- 229920001778 nylon Polymers 0.000 claims description 19
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 18
- 238000011534 incubation Methods 0.000 claims description 18
- 239000000463 material Substances 0.000 claims description 15
- 210000002966 serum Anatomy 0.000 claims description 15
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 12
- 229940098773 bovine serum albumin Drugs 0.000 claims description 12
- 238000001514 detection method Methods 0.000 claims description 12
- 229920000136 polysorbate Polymers 0.000 claims description 12
- 239000000758 substrate Substances 0.000 claims description 12
- 239000007790 solid phase Substances 0.000 claims description 10
- 238000000018 DNA microarray Methods 0.000 claims description 7
- 239000011159 matrix material Substances 0.000 claims description 6
- 238000012216 screening Methods 0.000 claims description 6
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- 239000003814 drug Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
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- 238000004519 manufacturing process Methods 0.000 claims description 3
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- 150000003839 salts Chemical class 0.000 claims description 3
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- 238000000034 method Methods 0.000 abstract description 14
- 230000000903 blocking effect Effects 0.000 abstract 3
- FKNQFGJONOIPTF-UHFFFAOYSA-N Sodium cation Chemical class [Na+] FKNQFGJONOIPTF-UHFFFAOYSA-N 0.000 abstract 1
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- YMXHPSHLTSZXKH-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 5-(2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl)pentanoate Chemical compound S1CC2NC(=O)NC2C1CCCCC(=O)ON1C(=O)CCC1=O YMXHPSHLTSZXKH-UHFFFAOYSA-N 0.000 description 1
- YMXHPSHLTSZXKH-RVBZMBCESA-N (2,5-dioxopyrrolidin-1-yl) 5-[(3as,4s,6ar)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoate Chemical compound C([C@H]1[C@H]2NC(=O)N[C@H]2CS1)CCCC(=O)ON1C(=O)CCC1=O YMXHPSHLTSZXKH-RVBZMBCESA-N 0.000 description 1
- HKAVADYDPYUPRD-UHFFFAOYSA-N 1h-pyrazine-2-thione Chemical compound SC1=CN=CC=N1 HKAVADYDPYUPRD-UHFFFAOYSA-N 0.000 description 1
- LLIANSAISVOLHR-GBCQHVBFSA-N 5-[(3as,4s,6ar)-2-oxidanylidene-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoic acid Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21.N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 LLIANSAISVOLHR-GBCQHVBFSA-N 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
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- 208000016610 ataxia-hypogonadism-choroidal dystrophy syndrome Diseases 0.000 description 1
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- 229940042795 hydrazides for tuberculosis treatment Drugs 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
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- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention relates to a protein chip using a label streptavidin-biotin technique. A sample is applied to a protein to be made. The hybridization reaction is carried out by the following steps that a sample to tested is dripped on the chip where the sample is applied; redundant samples are washed by washing liquid mainly containing sodium ion salt with low concentration; blocking liquid is added to block the chip, and the chip is cleaned again and dried; the biotin labeled protein diluted with the blocking liquid is dripped, and the chip is cleaned and dried at room temperature; fluorescence or enzymes labeled streptavidin diluted with the blocking liquid are dripped, and the chip is cleaned and dried at room temperature. The protein chip technique is used for testing antigens and antibodies, and is used for testing substances specifically combined with the protein. The protein chip improves a making method, the test result is clear and stable, and the reliability is strong. The protein chip uses different labeling methods to make labels on proteins with different properties; thus, the measurable protein varieties are increased, and the application range of the protein chip is widened.
Description
Technical field
The invention belongs to life science, particularly a kind of employing fluorescence or enzyme labeling Streptavidin-biotin technology (Fluorescence/Enzyme labeled Streptavidin Biotin Method, protein-chip FLSAB/ELSAB).
Background technology
Labelled streptavidin-biotin technology is a development in recent years new bio reaction rapidly amplification system, can be applicable to immunoassays.
The protein-chip of utilization routine immunization labelling technique, fluorescence or enzyme directly be marked at can with protein that the test substance specificity combines on, through further amplifying, detection sensitivity does not have limitation to reaction result, and equipment, reagent and personnel operation etc. are had relatively high expectations.Because some nonspecific combinations, background has some undesired signals in the course of reaction, under or the situation that condition control is poor slightly higher in point sample concentration, also false positive reaction may occur.In addition, because fluorescent marker is difficult to be marked on the acidic protein, its utilization scope is subjected to certain restriction, and for example: some antigen is acidic protein, and the efficient of fluorescent marker labelled antigen is extremely low, can't directly detect antibody with fluorescent labeled antigen.And the enzyme mark can influence the joint efficiency of antigen-antibody owing to the steric effect of molecule.
Summary of the invention
The purpose of this invention is to provide a kind of protein-chip of employing labelled streptavidin-biotin technology, it has improved method for making, makes testing result clear, stable, good reliability.
Another purpose of the present invention provides the application process of the protein-chip of employing labelled streptavidin-biotin technology, utilizes different labeling methods to mark on protein of different nature, enlarges the range of application of protein-chip.
Technical scheme of the present invention is as follows:
A kind of protein-chip of employing labelled streptavidin-biotin technology, the point sample and the manufacturing technology of protein are taked the following step:
(1) use the high speed spotting robot that protein example antigen or antibody point are printed on through on the microslide that glutaraldehyde was handled or on nitrocellulose filter or the nylon membrane;
(2) be fixed in 1 hour on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled in ambient temperature overnight or in 37 ℃ of incubations;
(3) add confining liquid on the microslide of handling through glutaraldehyde or nitrocellulose filter or nylon membrane surface, in 37 ℃ of incubations 1 hour based on bovine serum albumin(BSA) and tween;
(4) fully wash and drying at room temperature with distilled water;
Hybridization reaction is taked the following step:
(1) drips testing sample at point on the excellent chip,, antigen-antibody is fully reacted in 37 ℃ of incubations 1 hour;
(2) be the main unnecessary sample of eluent flush away with low concentration sodion salt, dry in room temperature;
(3) in addition bovine serum albumin(BSA) and tween are that main confining liquid is blockaded and cleaned once more and dry;
(4) drip the biotin labeled protein that the confining liquid based on bovine serum albumin(BSA) and tween diluted, in 37 ℃ of incubations 30 minutes;
(5) clean and dry in room temperature;
(6) drip fluorescence or the enzyme labeling Streptavidin that the confining liquid based on bovine serum albumin(BSA) and tween diluted, in 37 ℃ of lucifuge incubations 30 minutes;
(7) clean and dry in room temperature;
(8) if adopt the enzyme labeling Streptavidin in (6), then drip substrate, lucifuge incubation 30 minutes is with the stop buffer cessation reaction;
Reaction utilizes the chip scanning analyser that the result is carried out interpretation, testing result after finishing.
Described protein biochip technology is used to measure antigen, will with corresponding various monoclonal antibodies of determined antigen or polyclonal antibody dot matrix, be fixed on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled, antibody that these are excessive and patients serum's antigen combination, behind unconjugated other material of flush away, the Streptavidin that adds biotin labeled another antibody-fluorescence or enzyme labeling, after washing solid phase once more, measure fluorescence intensity, perhaps add the relative substrate of enzyme, and the gray-scale value after the survey change color, its value is directly proportional with tested antigen concentration.
Described protein biochip technology is used to measure antibody, will be with the corresponding antigen dot matrix of antibody to be measured, be fixed on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled, antigen that these are excessive and patients serum's antibodies, behind unconjugated other material of flush away, the Streptavidin that adds biotin labeled another antigen or two anti--fluorescence or enzyme labeling, after washing solid phase once more, measure fluorescence intensity, perhaps add the relative substrate of enzyme, and the gray-scale value after the survey change color, its value is directly proportional with tested antibody concentration.
Described protein biochip technology is used to detect the material that combines with protein specific, will with the corresponding protein array of test substance, be fixed on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled, these excess protein and test substance combination, behind unconjugated other material of flush away, add biotin labeled another the protein-fluorescence that combines with test substance or the Streptavidin of enzyme labeling, after washing solid phase once more, measure fluorescence intensity, perhaps add the relative substrate of enzyme, and the gray-scale value after the survey change color, its value is directly proportional with tested material concentration, this detection method is used for part, the detection of acceptor, screening also is used for the research of signal conduction and the screening of medicine.
The protein-chip of employing fluorescence of the present invention or enzyme labeling Streptavidin-biotin technology, compare with the protein-chip that adopts the routine immunization fluorescent technique, reaction reagent be with biotin (Biotin) but the specificity of mark in conjunction with the protein (as antibody) of test substance (as antigen), with the Streptavidin (Streptavidin) of fluorescence (Fluorescence) or enzyme (Enzyme) mark, utilize the specificity high affinity of Streptavidin and biotin to carry out the biological respinse amplification.
Protein-chip of the present invention has following advantage:
1, but sensitivity and sensing range improve more than 10 times than the protein-chip that adopts the conventional tag method, and its detection signal is strong.
2, have high degree of specificity, background signal is low, and it is non-specific painted light, and testing result is clear, stable, good reliability.
But the sample high dilution further reduces the painted possibility of background when 3, operating, and background noise is low, and specificity further improves.
4, acted on the high efficiency of the little array technology of protein-chip, easy and simple to handle.
5, utilize different labeling methods to mark on protein of different nature, the kind of detectable protein increases thus, enlarges the range of application of protein-chip.
6, reduce the steric effect of enzyme, improve reaction efficiency.
The present invention utilizes different biotin labeling methods, can mark protein of different nature, enlarge the range of application of protein-chip.For example, (biotinyl-N-hydroxy-succinimide ester BNHS) is applicable to the protein of mark neutrality or meta-alkalescence for biotin N-maloyl imines ester with biotin activation; (biotin hydrazide BHZ) is mainly used in the glycoprotein of mark slant acidity for the biotin hydrazides with biotin activation.
Description of drawings
Below in conjunction with drawings and Examples the present invention is elaborated.
Fig. 1 is the schematic diagram that adopts the protein-chip mensuration antibody (Ag-Ab-two resistive connections close) of fluorescence labeling Streptavidin-biotin technology.
Fig. 2 is the schematic diagram that adopts the protein-chip mensuration antigen (double antibody sandwich method) of fluorescence labeling Streptavidin-biotin technology.
Fig. 3 is the schematic diagram that adopts the protein-chip mensuration antibody (dual-antigen sandwich method) of fluorescence labeling Streptavidin-biotin technology.
Fig. 4 is the schematic diagram that adopts the protein-chip mensuration antibody (Ag-Ab-two resistive connections close) of enzyme labeling Streptavidin-biotin technology.
Fig. 5 is the schematic diagram that adopts the protein-chip mensuration antigen (double antibody sandwich method) of enzyme labeling Streptavidin-biotin technology.
Fig. 6 is the schematic diagram that adopts the protein-chip mensuration antibody (dual-antigen sandwich method) of enzyme labeling Streptavidin-biotin technology.
Fig. 7 is the fluorescent scanning result schematic diagram that adopts the protein-chip detection patients serum of fluorescence labeling Streptavidin-biotin technology.
Fig. 8 is to use the fluorescent scanning result schematic diagram of single fluorescence labeling technology for detection original content serum.
Fig. 9 adopts the protein-chip of fluorescence labeling Streptavidin-biotin technology to detect the patients serum, and the patients serum is diluted experimental result synoptic diagram after 10 times.
Figure 10 is to use single fluorescence labeling technology for detection original content serum, and the patients serum is diluted experimental result synoptic diagram after 10 times.
Embodiment
The present invention is a kind of protein-chip of employing labelled streptavidin-biotin technology, and the solid phase carrier of this protein-chip can be microslide or nitrocellulose filter (NC film) or nylon membrane.
The point sample and the manufacturing technology of protein are taked the following step:
(1) use the high speed spotting robot that protein example antigen or antibody point are printed on through on the microslide that glutaraldehyde was handled or on nitrocellulose filter or the nylon membrane.
(2) be fixed in 1 hour on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled in ambient temperature overnight or in 37 ℃ of incubations.
(3) add confining liquid on the microslide of handling through glutaraldehyde or nitrocellulose filter or nylon membrane surface, in 37 ℃ of incubations 1 hour based on bovine serum albumin(BSA) (BSA) and tween.
(4) fully wash and drying at room temperature with distilled water.
Hybridization reaction is taked the following step:
(1) drips testing sample at point on the excellent chip,, antigen-antibody is fully reacted in 37 ℃ of incubations 1 hour.
(2) be the main unnecessary sample of eluent flush away with low concentration sodion salt, dry in room temperature.
(3) in addition bovine serum albumin(BSA) and tween are that main confining liquid is blockaded and cleaned once more and dry.
(4) drip the biotin labeled protein that the confining liquid based on bovine serum albumin(BSA) and tween diluted, in 37 ℃ of incubations 30 minutes.
(5) clean and dry in room temperature.
(6) drip fluorescence or the enzyme labeling Streptavidin that the confining liquid based on bovine serum albumin(BSA) and tween diluted, in 37 ℃ of lucifuge incubations 30 minutes.
(7) clean and dry in room temperature.
(8) if adopt the enzyme labeling Streptavidin in (6), then drip substrate, lucifuge incubation 30 minutes is with the stop buffer cessation reaction.
After reaction finishes, utilize the chip scanning analyser of specialty that the result is carried out interpretation, testing result.
Application process of the present invention is as follows:
Referring to Fig. 1, Fig. 3, Fig. 4 and Fig. 6, protein biochip technology is used to measure antibody, will with the corresponding antigen dot matrix of antibody to be measured, be fixed on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled, antigen that these are excessive and patients serum's antibodies, behind unconjugated other material of flush away, the Streptavidin that adds biotin labeled another antigen or two anti--fluorescence or enzyme labeling, after washing solid phase once more, measure fluorescence intensity, perhaps add the relative substrate of enzyme, and the gray-scale value after the survey change color, its value is directly proportional with tested antibody concentration.
Referring to Fig. 2 and Fig. 4, protein-chip is used to measure antigen, will with corresponding various monoclonal antibodies of determined antigen or polyclonal antibody dot matrix, be fixed on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled, antibody that these are excessive and patients serum's antigen combination, behind unconjugated other material of flush away, the Streptavidin that adds biotin labeled another antibody-fluorescence or enzyme labeling, after washing solid phase once more, measure fluorescence intensity, perhaps add the relative substrate of enzyme, and the gray-scale value after the survey change color, its value is directly proportional with tested antigen concentration.
Protein-chip is used to detect the material that combines with protein specific, will with the corresponding protein array of test substance, be fixed on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled, these excess protein and test substance combination, behind unconjugated other material of flush away, add biotin labeled another the protein-fluorescence that combines with test substance or the Streptavidin of enzyme labeling, after washing solid phase once more, measure fluorescence intensity, perhaps add the relative substrate of enzyme, and the gray-scale value after the survey change color, its value is directly proportional with tested material concentration, this detection method is used for part, the detection of acceptor, screening also is used for the research of signal conduction and the screening of medicine.
The present invention is one of them embodiment with the detection of third liver clinical examination index hepatitis C virus (HCV) commonly used:
Referring to Fig. 7 to Figure 10, detect the resulting fluorescent scanning result of certain patients serum with protein-chip, 10 points of first row and second row are testing result among the figure, last two behavior positive controls, the negative contrast of middle two rows no signal.
Fig. 7 and Fig. 8 are for detecting original content serum, and wherein Fig. 7 has utilized fluorescence labeling Streptavidin-biotin technology, and Fig. 8 then uses single fluorescence labeling technology.As can be seen from the figure, adopt the reaction result positive reaction signal of fluorescence labeling Streptavidin-biotin technology strong, clear picture, background signal is low; And use the reaction result of single fluorescence labeling technology, and background signal is higher relatively, and image is fuzzy relatively.
Fig. 9 and Figure 10 dilute experimental result after 10 times with the patients serum.Fig. 9 has utilized fluorescence labeling Streptavidin-biotin technology, and Figure 10 then uses single fluorescence labeling technology, and as seen the former result is still very clear, and latter's consequential signal is very weak, and image blurring difficulty is debated, and may produce false-negative conclusion thus.
Claims (4)
1. protein-chip that adopts labelled streptavidin-biotin technology, the point sample and the manufacturing technology of protein are taked the following step:
(1) use the high speed spotting robot that protein example antigen or antibody point are printed on through on the microslide that glutaraldehyde was handled or on nitrocellulose filter or the nylon membrane;
(2) be fixed in 1 hour on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled in ambient temperature overnight or in 37 ℃ of incubations;
(3) add the confining liquid that contains bovine serum albumin(BSA) and tween at the microslide of handling through glutaraldehyde or nitrocellulose filter or nylon membrane surface, in 37 ℃ of incubations 1 hour;
(4) fully wash and drying at room temperature with distilled water;
Hybridization reaction is taked the following step:
(1) drips testing sample at point on the excellent chip,, antigen-antibody is fully reacted in 37 ℃ of incubations 1 hour;
(2) with the unnecessary sample of eluent flush away that contains low concentration sodion salt, dry in room temperature;
(3) add that the confining liquid that contains bovine serum albumin(BSA) and tween is blockaded and clean once more and dry;
It is characterized in that hybridization reaction is then taked the following step:
(4) drip the biotin labeled protein that the confining liquid contain bovine serum albumin(BSA) and tween diluted, in 37 ℃ of incubations 30 minutes;
(5) clean and dry in room temperature;
(6) drip fluorescence or the enzyme labeling Streptavidin that the confining liquid contain bovine serum albumin(BSA) and tween diluted, in 37 ℃ of lucifuge incubations 30 minutes;
(7) clean and dry in room temperature;
(8) if adopt the enzyme labeling Streptavidin in (6), then drip substrate, lucifuge incubation 30 minutes is with the stop buffer cessation reaction;
Reaction utilizes the chip scanning analyser that the result is carried out interpretation, testing result after finishing.
2. the protein-chip of employing labelled streptavidin-biotin technology according to claim 1, it is characterized in that, described protein biochip technology is used to measure antigen, will with corresponding various monoclonal antibodies of determined antigen or polyclonal antibody dot matrix, be fixed on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled, antibody that these are excessive and patients serum's antigen combination, behind unconjugated other material of flush away, add biotin labeled in addition-Streptavidin of antibody-fluorescence or enzyme labeling, after washing solid phase once more, measure fluorescence intensity, perhaps add the relative substrate of enzyme, and the gray-scale value after the survey change color, its value is directly proportional with tested antigen concentration.
3. the protein-chip of employing labelled streptavidin-biotin technology according to claim 1, it is characterized in that, described protein biochip technology is used to measure antibody, will with the corresponding antigen dot matrix of antibody to be measured, be fixed on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled, antigen that these are excessive and patients serum's antibodies, behind unconjugated other material of flush away, the Streptavidin that adds biotin labeled another antigen or two anti--fluorescence or enzyme labeling, after washing solid phase once more, measure fluorescence intensity, perhaps add the relative substrate of enzyme, and the gray-scale value after the survey change color, its value is directly proportional with tested antibody concentration.
4. the protein-chip of employing labelled streptavidin-biotin technology according to claim 1, it is characterized in that, described protein biochip technology is used to detect the material that combines with protein specific, will with the corresponding protein array of test substance, be fixed on microslide or nitrocellulose filter or nylon membrane that glutaraldehyde was handled, these excess protein and test substance combination, behind unconjugated other material of flush away, add biotin labeled another the protein-fluorescence that combines with test substance or the Streptavidin of enzyme labeling, after washing solid phase once more, measure fluorescence intensity, perhaps add the relative substrate of enzyme, and the gray-scale value after the survey change color, its value is directly proportional with tested material concentration, this detection method is used for part, the detection of acceptor, screening also is used for the research of signal conduction and the screening of medicine.
Priority Applications (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011133236A CN1156702C (en) | 2001-07-11 | 2001-07-11 | Protein chip based on labeling streptavidin-biotin technology |
| JP2002576595A JP2004532404A (en) | 2001-03-28 | 2002-03-28 | Apparatus and method for detecting multiple analytes |
| EP02721953A EP1373467A4 (en) | 2001-03-28 | 2002-03-28 | Device and method for detection of multiple analytes |
| CNA028075285A CN1500140A (en) | 2001-03-28 | 2002-03-28 | Device and method for measuring a great variety of matters to be analyzed |
| US10/472,806 US20040091939A1 (en) | 2001-03-28 | 2002-03-28 | Device and method for detection of multiple analytes |
| PCT/CN2002/000209 WO2002077152A1 (en) | 2001-03-28 | 2002-03-28 | Device and method for detection of multiple analytes |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011133236A CN1156702C (en) | 2001-07-11 | 2001-07-11 | Protein chip based on labeling streptavidin-biotin technology |
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| CN1335505A CN1335505A (en) | 2002-02-13 |
| CN1156702C true CN1156702C (en) | 2004-07-07 |
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| US7138262B1 (en) | 2000-08-18 | 2006-11-21 | Shire Human Genetic Therapies, Inc. | High mannose proteins and methods of making high mannose proteins |
| CN100338464C (en) * | 2002-12-27 | 2007-09-19 | 穆海东 | Protein chip carrier processing method |
| CN100341570C (en) * | 2003-03-31 | 2007-10-10 | 穆海东 | Preparation method of protein chip |
| JP2004309416A (en) * | 2003-04-10 | 2004-11-04 | Sony Corp | Sensor unit and sensing method, sensor unit and sensing method for biosubstance, sensor unit and sensing method for secrete, and feeling sensor unit and sensing method |
| PL1986612T3 (en) | 2006-02-07 | 2013-02-28 | Shire Human Genetic Therapies | Stabilized composition of glucocerebrosidase |
| NZ713967A (en) | 2009-07-28 | 2017-01-27 | Shire Human Genetic Therapies | Compositions and methods for treating gaucher disease |
| WO2012012461A2 (en) | 2010-07-19 | 2012-01-26 | Shire Human Genetic Therapies, Inc. | Mannose receptor c type 1 (mrc1) codon optimized cell line and uses thereof |
| MX361231B (en) * | 2012-03-02 | 2018-11-30 | Shire Human Genetic Therapies | Compositions and methods for treating type iii gaucher disease. |
| CN104569417B (en) * | 2013-10-12 | 2016-06-01 | 广州瑞博奥生物科技有限公司 | A kind of antibody chip test kit for early diagnosis acute injury of kidney |
| 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 |
| CN105510596A (en) * | 2015-12-02 | 2016-04-20 | 深圳市赛尔生物技术有限公司 | Protein chip for detecting respiratory virus antigens as well as kit and preparation method thereof |
| CN106198987A (en) * | 2016-08-31 | 2016-12-07 | 辽宁迈迪生物科技股份有限公司 | A kind of detection label for evaluating tumor vaccine cells therapy and detection method thereof and application |
| CN106754860A (en) * | 2016-11-22 | 2017-05-31 | 上海纳米技术及应用国家工程研究中心有限公司 | A kind of method to interface Rapid Modification Avidin based on liposome |
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