CN2489341Y - Directional fixed antibody protein chip - Google Patents
Directional fixed antibody protein chip Download PDFInfo
- Publication number
- CN2489341Y CN2489341Y CN 01233478 CN01233478U CN2489341Y CN 2489341 Y CN2489341 Y CN 2489341Y CN 01233478 CN01233478 CN 01233478 CN 01233478 U CN01233478 U CN 01233478U CN 2489341 Y CN2489341 Y CN 2489341Y
- Authority
- CN
- China
- Prior art keywords
- chip
- protein
- antibody
- albumin
- rete
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The utility model relates to a directional fixed antibody protein chip, comprising a solid base chip and an antibody inducting membrane layer. The utility model is characterized in that a protein A membrane layer is arranged between the solid base chip and the antibody inducting membrane layer. The utility model also comprises a lyophobicpolarization surface layer which is arranged between the solid base chip and the protein A membrane layer. the utility model has the advantages that the protein A has a function of arm connecting; and the antigen DBD of the antibody molecule extends out from the surface of the solid to combine with the antigen freely, thereby greatly increasing the conjugated antigen capability of the protein chip. The utility model is widely used for immunoassay, internal secretion detection and drug screening.
Description
Technical field
The utility model relates to the immune detection chip, particularly a kind of can be on solid surface the protein-chip of directional immobilizing antibody.
Background technology
Antibody molecule has high special selection natural quality to target antigen, it has been widely used on panimmunity diagnostic method and the panimmunity sensor, as T.M.Phillips etc. described in the document " Protein A coated glass beads universal support medium for high-performance immunoaffinity chromatography " (Journal of Chromatography, 327 (1985) 213-219).Wherein many is solid-phase immunity methods, antibody molecule is fixed on detects antigen on the solid surface exactly.The biologically active that directly is fixed on the antibody molecule on the solid phase surface is usually less than the antibody molecule in the solution, the main cause that biologically active reduces directly is fixed on the sterically hindered increase of the antibody molecule on the solid phase surface, be unfavorable for the combination of antibody-antigen, as document " Effectiveness of protein A for antibodyimmobilization for a fiber optic biosensor) " (Biosensors ﹠amp; Bioelectronics, Vo1.12 No.4,329-336,1997) described; The functional domain that the another one reason directly is fixed on the antibody molecule on the solid phase surface tends on solid phase surface downwards or during the trend side, is unfavorable for the combination of antibody-antigen; If also have the functional domain that combines with antigen on the antibody molecule to be adsorbed on the solid phase surface, antibody molecule just can not synantigen generation combination, as shown in Figure 1, antibody molecule 5 contains two antigen binding domains, one of them and solid phase surface 1 combine, thereby have lost the ability that combines with antigen molecule 6.
Summary of the invention
The purpose of this utility model is, overcoming the above-mentioned functional domain sterically hindered big, antibody molecule that directly is fixed on the antibody molecule on the solid phase surface is adsorbed on the solid phase surface at the functional domain downward or that tend to combine with antigen on side, the antibody molecule of trend on the solid phase surface, be unfavorable for the combination of antibody-antigen, in order to improve the antibody on the detection chip and the binding ability of antigen, thereby provide a kind of protein-chip of directional immobilizing antibody.
The purpose of this utility model is achieved in that
The protein-chip of the directional immobilizing antibody that the utility model provides comprises solid substrate and antibody sensor film, it is characterized in that: also have one deck albumin A rete between solid substrate and antibody sensor film.
Wherein said solid substrate is semiconductor material (as silicon chip, germanium wafer etc.), metal, glass, plastics or solid composite material (solid composite material as: semiconductor surface or glass surface metal-coated membrane).
The protein-chip of the directional immobilizing antibody that the utility model provides when solid substrate is semiconductor material, glass or plastics, also further comprises: also have a hydrophobic polarized meter surface layer between solid substrate and albumin A rete.This hydrophobic polarized meter surface layer is to adopt chemical treatment method to form, because form hydrophobic polarized meter surface layer on solid substrate, helps in the preparation process of albumin A rete adhesion protein A from solution, promotes the formation of albumin A rete.
Described hydrophobic polarized meter surface layer, albumin A rete, antibody sensor film are unimolecular layer.
Wherein said hydrophobic polarized meter surface layer can be the hydrophobic polarized meter surface layer of hydrophobic silicon alkane, as the hydrophobic polarized meter surface layer of dichlorodimethylsilane.
The protein-chip of directional immobilizing antibody provided by the invention uses albumin A to come the directional immobilizing antibody molecule.Albumin A is the protein that extracts from staphylococcus aureus, the Fc fragments specific combination that it can synantibody.This albumin A have five can specificity binding antibody molecule Fc fragment functional domain (E, D, A, B and C), each functional domain contains 58 amino acid residues, and each functional domain can both the combination of independent same Fc fragment.Albumin A is the Fc fragment of binding antibody molecule specifically, and it is outer so that the combination of synantigen molecule to make on the antibody molecule two functional domain Fab of synantigen molecular specificity combination stretch to the surface.As shown in Figure 2, directed two functional domains that are fixed on antibody molecule 5 synantigen molecules 6 combinations of solid surface 1 stretch to outside the surface synantigen molecule 6 combinations freely by the combination of albumin A 3 specificitys.The albumin A that is combined on the solid surface plays the effect that arm connects, and connected antibody molecule is fully stretched in solution, has greatly reduced space steric effect, makes the association class between antibody-antigen be similar in solution equally free.In addition, the albumin A rete can also the passivation solid surface, reduces the non-specific adsorption of solid surface effectively, improves the sensitivity for analysis that immune detection promptly detects antigen.
The antibody of antibody sensor film of the present invention comprises the various antibody that are used for immune detection, as immunoglobulin G while antibody (antiIgG), bovine serum albumin(BSA) antibody (antiBSA) or human serum blood coagulating protein original antibody (antiFIB) etc.
The protein-chip of the directional immobilizing antibody that the utility model provides can also be used for the research of subjects such as life science, particularly immunology, molecular biology, biological chemistry, oncology, cell biology except that carrying out biological detection.
Advantage of the present utility model and effect:
1. the protein-chip of the directional immobilizing antibody that provides of the utility model has overcome on the solid surface shortcoming that antibody biologically active that direct sessile antibody causes descends;
2. protein-chip of the present utility model, directed fixing antibody molecule makes the antigen binding domain on the antibody molecule stretch to outside the solid surface synantigen combination freely;
3. protein-chip of the present utility model, albumin A have wherein played the effect that arm connects, and have reduced the sterically hindered of antibody molecule;
4. the albumin A rete can also passivated surface, reduces the non-specific adsorption on surface effectively, the check and analysis sensitivity that improves Detection of antigen;
5. the regeneration of chip albumin A rete is simple, and chip can be reused repeatedly.
Description of drawings
Fig. 1 is the antibody molecule synoptic diagram that directly is fixed on the solid phase surface;
Fig. 2 is the fixing antibody molecule synoptic diagram of orientation of the present invention;
Fig. 3 is the protein-chip structural representation of the directional immobilizing antibody of the utility model embodiment 3;
Wherein
1: solid substrate 2: hydrophobic polarized meter surface layer 3: albumin A rete
4: antibody sensor film 5: antibody molecule 6: antigen molecule
Embodiment
The protein-chip of directional immobilizing antibody immunoglobulin G while (antiIgG) on the semi-conductor silicon chip surface, this protein-chip structure comprises: the silicon chip 1 that the thick integrated circuit of a 0.5mm is used, the hydrophobic polarized meter surface layer 2 of unimolecular layer dichlorodimethylsilane that on silicon chip 1, forms, the unimolecular layer albumin A rete 3 that on hydrophobic polarized meter surface layer 2, forms, and the unimolecular layer antiIgG sensor film 4 that on albumin A rete 3, forms.
The protein-chip of directional immobilizing antibody bovine serum albumin(BSA) (antiBSA) on the gold-plated composite substrate of silicon chip surface, this protein-chip structure comprises: be coated with the complex solid substrate 1 that the thick gold plaque of 2 μ m forms on the silicon chip that the thick integrated circuit of a 0.5mm is used, the unimolecular layer albumin A rete 3 that on complex solid substrate 1 gold plaque, forms, and the unimolecular layer antiBSA sensor film 4 that an end forms to 2/3 part of other end entire area on albumin A rete 3.
The directed fixedly protein-chip of human serum blood coagulating protein original antibody (antiFIB) on silicon chip surface, this protein-chip structure as shown in Figure 3, comprise: the silicon chip 1 that the thick integrated circuit of a 0.5mm is used, the hydrophobic polarized meter surface layer 2 of unimolecular layer dichlorodimethylsilane that on silicon chip 1, forms, the unimolecular layer albumin A rete 3 that on hydrophobic polarized meter surface layer 2, forms, and the unimolecular layer antiFIB sensor film 4 that an end to 1/2 part of other end entire area forms on albumin A rete 3.
The orientation of present embodiment fixedly 1/2 part of the protein-chip sensor film end of antiFIB be immersed in the solution to be measured, if contain the FIB molecule in the solution, just together with the antibody generation specificity combination on the chip, form compound molecule, cause surface density (or thicknesses of layers) to increase, otherwise surface density (or thicknesses of layers) just can not change.Whether rete changes can be arrived by the ellipsometry imaging systematic observation, judges that with this whether the FIB molecule exists in solution.
Embodiment 4
The protein-chip of directional immobilizing antibody immunoglobulin G while (antiIgG) on frosting, this protein-chip structure comprises: the plastic sheet 1 that a 0.8mm is thick, the hydrophobic polarized meter surface layer 2 of unimolecular layer dichlorodimethylsilane that on plastic sheet 1, forms, the unimolecular layer albumin A rete 3 that on hydrophobic polarized meter surface layer 2, forms, and the unimolecular layer antiIgG sensor film 4 that on albumin A rete 3, forms.
Claims (9)
1. the protein-chip of a directional immobilizing antibody comprises solid substrate and antibody sensor film, it is characterized in that: also have one deck albumin A rete between solid substrate and antibody sensor film.
2. the protein-chip of directional immobilizing antibody according to claim 1 is characterized in that: also comprise the hydrophobic polarized meter surface layer of one deck between solid substrate and albumin A rete.
3. the protein-chip of directional immobilizing antibody according to claim 1, it is characterized in that: described solid substrate is the solid composite material of metal or plating metal on surface film.
4. the protein-chip of directional immobilizing antibody according to claim 2, it is characterized in that: described solid substrate is semiconductor material silicon chip, semiconductor material germanium wafer, metal, glass, plastics or solid composite material.
5. according to the protein-chip of claim 3 or 4 described directional immobilizing antibodies, it is characterized in that: described solid composite material is the silicon chip of plating metal on surface film, the germanium wafer of plating metal on surface film, the plastic sheet of plating metal on surface film or the glass of plating metal on surface film.
6. the protein-chip of directional immobilizing antibody according to claim 1, it is characterized in that: described albumin A rete and antibody sensor film are unimolecular layer.
7. the protein-chip of directional immobilizing antibody according to claim 2, it is characterized in that: described hydrophobic polarized meter surface layer, albumin A rete and antibody sensor film are unimolecular layer.
8. the protein-chip of directional immobilizing antibody according to claim 2, it is characterized in that: the antibody of described antibody sensor film comprises the various antibody molecules that are used for immune detection.
9. the protein-chip of directional immobilizing antibody according to claim 2, it is characterized in that: described hydrophobic polarized meter surface layer is the hydrophobic polarized meter surface layer of hydrophobic silane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01233478 CN2489341Y (en) | 2001-08-20 | 2001-08-20 | Directional fixed antibody protein chip |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01233478 CN2489341Y (en) | 2001-08-20 | 2001-08-20 | Directional fixed antibody protein chip |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN2489341Y true CN2489341Y (en) | 2002-05-01 |
Family
ID=33647301
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 01233478 Expired - Fee Related CN2489341Y (en) | 2001-08-20 | 2001-08-20 | Directional fixed antibody protein chip |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN2489341Y (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103323592A (en) * | 2012-03-22 | 2013-09-25 | 曾立波 | High-throughput drug rapid detection chip and system |
| CN103558376A (en) * | 2013-10-14 | 2014-02-05 | 青岛海佑海洋生物工程有限公司 | Novel polysaccharide biological electrode chip and preparation method thereof |
| CN103728447A (en) * | 2014-01-03 | 2014-04-16 | 北京晶泰美康生物科技有限公司 | Controllable quantum dot locus specificity bridging coupling antibody marking method and application |
| CN106771143A (en) * | 2016-11-30 | 2017-05-31 | 天津大学 | The preparation method of the prostate cancer antigen immuno-chromatographic test paper strip of low concentration antibody is efficiently fixed based on hydrophobin |
-
2001
- 2001-08-20 CN CN 01233478 patent/CN2489341Y/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103323592A (en) * | 2012-03-22 | 2013-09-25 | 曾立波 | High-throughput drug rapid detection chip and system |
| CN103323592B (en) * | 2012-03-22 | 2015-09-23 | 曾立波 | High-throughput drug rapid detection chip and system |
| CN103558376A (en) * | 2013-10-14 | 2014-02-05 | 青岛海佑海洋生物工程有限公司 | Novel polysaccharide biological electrode chip and preparation method thereof |
| CN103728447A (en) * | 2014-01-03 | 2014-04-16 | 北京晶泰美康生物科技有限公司 | Controllable quantum dot locus specificity bridging coupling antibody marking method and application |
| CN103728447B (en) * | 2014-01-03 | 2016-07-13 | 北京纳晶生物科技有限公司 | The antibody labeling method of controllable quantum dots locus specificity bridge joint coupling and application thereof |
| CN106771143A (en) * | 2016-11-30 | 2017-05-31 | 天津大学 | The preparation method of the prostate cancer antigen immuno-chromatographic test paper strip of low concentration antibody is efficiently fixed based on hydrophobin |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10585092B2 (en) | Devices and methods for the rapid and accurate detection of analytes | |
| JP4234596B2 (en) | Method for analysis and biosensor | |
| Makaraviciute et al. | Site-directed antibody immobilization techniques for immunosensors | |
| Lee et al. | Fabrication of self-assembled protein A monolayer and its application as an immunosensor | |
| Jang et al. | Modified fabrication process of protein chips using a short-chain self-assembled monolayer | |
| Peluso et al. | Optimizing antibody immobilization strategies for the construction of protein microarrays | |
| JP3865409B2 (en) | Affinity array with a wide range of specificities: a quantitative method for the identification of complex samples | |
| Melnyk et al. | Peptide arrays for highly sensitive and specific antibody-binding fluorescence assays | |
| Bashir et al. | Adsorption of avidin on microfabricated surfaces for protein biochip applications | |
| JP4443764B2 (en) | Method for immobilizing biological molecules on a support surface | |
| JP2008082988A (en) | Detecting method utilizing multistage amplification | |
| Chandra et al. | Prospects and advancements in C-reactive protein detection | |
| CN2489341Y (en) | Directional fixed antibody protein chip | |
| JP4768417B2 (en) | Biosensor | |
| TW201124721A (en) | Molecular probe chip with covalent bonding organoconductive anchoring compound | |
| JP2005037331A (en) | Substrate for detecting substance derived from organism and its manufacturing method | |
| JPS5916669B2 (en) | Antigen or antibody detection method | |
| JPH05506314A (en) | Method of manufacturing test surfaces for optical detection of biological binding | |
| CN1402007A (en) | Protein chip of directional immobilizing antibody, its preparing method and use thereof | |
| US8153445B2 (en) | Method for screening of infectious agents in blood | |
| Chiu et al. | Highly sensitive carboxyl-graphene oxide-based SPR immunosensor for the detection of CA19-9 biomarker | |
| JP5205293B2 (en) | Antibody-immobilized substrate, and method and use of the antibody-immobilized substrate | |
| CN1482456A (en) | Micro-array protein chip and making method thereof | |
| Demirel et al. | Oriented immobilization of IgG on hydroxylated Si (001) surfaces via protein-A by a multiple-step process based on a self-assembly approach | |
| CA2397500A1 (en) | Method for immobilisation of (an) affinity reagent(s) on a hydrophobic solid phase |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20020501 Termination date: 20100820 |