CN200989904Y - Polypeptide biochip structure for simulating acceptor signal molecule - Google Patents
Polypeptide biochip structure for simulating acceptor signal molecule Download PDFInfo
- Publication number
- CN200989904Y CN200989904Y CN 200620048746 CN200620048746U CN200989904Y CN 200989904 Y CN200989904 Y CN 200989904Y CN 200620048746 CN200620048746 CN 200620048746 CN 200620048746 U CN200620048746 U CN 200620048746U CN 200989904 Y CN200989904 Y CN 200989904Y
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- sephadex
- layer
- linking arm
- biochip
- 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 - Lifetime
Links
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The utility model relates to a polypeptide biochip structure simulating a receptor signal molecular; the utility model is characterized in that the polypeptide chip is a gilded surface membrane (2) on a glass bottom layer (1); a glucan gel layer (3) which is connected with a 6-amido caproic acid as a connecting arm (10) is fixed on the gilded surface membrane (2); the other end of the connecting arm (10) is connected with a phosphoric acid gene on a polypeptide molecular tyrosine residue; the polypeptide of the connecting arm is fixed with the glucan gel layer by the fixing method of standard amidogen. The utility model has the benefits of that a fast, real-time of technical platform without separation and labeling is built for the interaction of signal transduction protein and polypeptide drug filtration which takes the receptor as a target.
Description
Technical field
The utility model is a kind of polypeptide biochip structure of simulated receptor signaling molecule, or rather, the structure that relates to a kind of polypeptide biochip based on the surface plasma biology sensor is mainly used in the interaction dynamics of albumen action site peptide sequence and its associated protein in the transduction of observation receptor signal.Belong to the biochip field.
Background technology
Surface plasma body resonant vibration (SPR) biology sensor has been used for the biological and biomedical various aspects of research, comprises DNA-albumen, RNA-albumen, protein-protein, enzyme-substrate and polypeptide-albumen, the interaction between polypeptide-polypeptide.Wherein protein-protein polypeptide-protein-interacting classic method research covalent coupling or radioactivity of adopting are competed combination etc. more, and, the recombinant clone of protein and expression are very time-consuming and expensive, and study polypeptide and protein interaction with surface plasmon resonance biosensor, and the effect research that the domain that adopts polypeptide to constitute substitutes complete albumen has fast, in real time, need not advantages such as separation and mark.The key of utilization surface plasmon resonance biosensor is the preparation of biochip chip.
Summary of the invention
The utility model has been described a kind of polypeptide biochip structure of simulated receptor signaling molecule, and the domain (motif) that constitutes with peptide sequence replaces whole albumen, the preparation polypeptide chip.Can be used for the interactional measurement of receptor protein.
Described polypeptide biochip structure is to plate metal facial mask 2 on glass bottom 1, fixing sephadex layer 3 on it, be connected with a 6-aminocaprolc acid on the peptide molecule as linking arm 10 at sephadex layer 3, the linking arm other end connects the phosphoric acid gene 11 on the peptide molecule tyrosine residue.Linking arm 10 by standard amino with the sephadex molecular layer on the hydroxyl covalent coupling.
Usefulness of the present utility model is, for signal transducer interact and acceptor be the polypeptide drugs screening of target spot build fast, in real time, need not to separate and need not the technology platform of mark.
Description of drawings
Fig. 1 is the variation (b) that surface plasma resonance biosensor schematic diagram (a) and resonance signal take place.
Fig. 2 is based on the polypeptide biochip structure of the simulated receptor signaling molecule of surface plasmon resonance biosensor.
Fig. 3 is a kind of peptide sequence.Aha is 6-aminocaprolc acid (NH
2(CH
2)
6COOH), pY is a tyrosine phosphatase, NH
2Be the c-terminus aminoacylation, the sequence name of this polypeptide is called: the amino hexanoyl leucyl of 6-tyrosyl alanyl seryl valyl aspartoyl glutamy prolyl phosphoric acid tyrosyl ammonia
1. glass-based bottoms among the figure, 2. golden watch facial mask, 3. sephadex layer, 4. subdue ripple, 5. sample liquid, 6. prism, 7. incident polarized light, 8. incident angle, reflect polarized light 10. linking arms 11. phosphoric acid genes after 9. reflect polarized light and reflected light deviation angle change with the refractive index of decaying wave
Embodiment
Below by the introduction of specific embodiment, further illustrate characteristics of the present utility model and progress.
See also the polypeptide biochip structure of a kind of simulated receptor signaling molecule that Fig. 2, Fig. 2 provide for the utility model, plated with gold surface film 2 on glass-based bottom 1, fixing sephadex layer 3 on it connects by the hydroxyl covalent coupling on aminoterminal and the sephadex molecule as an end of linking arm 10; 6-aminocaprolc acid on peptide molecule is as linking arm, and the other end of linking arm 10 is to link to each other with the sephadex layer.Glass bottom 1, golden watch facial mask 2 and sephadex layer 3 are chip surface three-deckers from the incident light plane to the flow chamber.Signal protein interaction sites related polypeptide motif sequence in design and the definite signal transduction, synthetic c-terminus amidation and amido end add the polypeptide of 6-aminocaprolc acid as linking arm, utilize the amino fixing means of standard to be fixed in the sephadex layer, constitute the polypeptide biochip.The interaction protein of variable concentrations can be measured associated protein and the interactional kinetic constant of polypeptide in the receptor signal path in real time by this polypeptide chip surface, can be used for polypeptide drugs screening in the signal transduction.
Mimic peptide with a kind of para-insulin receptor protein and signaling molecule thereof is the concrete embodiment of example explanation.Query protein database and pertinent literature, find para-insulin receptor protein etc. to need to be difficult for obtaining respective action site polypeptide motif (motif) in the research two interaction albumen, add that at aminoterminal linking arm a: Aha (6-carbohexamide) is particularly when motif determines amino acid residue at c-terminus (as SH2 domain effect motif).The 1-5mg/ml polypeptide is dissolved in the HBS-EP damping fluid that contains 1-3mol/L NaCl, adopt the amino fixing means (seeing www.biacore.com) of BIAcore standard, be that the 5ul/min polypeptide solution is by F1 passage 7min, polypeptide fixedly desired concn reduces and corresponding increasing with polypeptide pI value, generally when polypeptide pI＜3.5, need peptide concentration 5mg/ml, when polypeptide pI＞6.0, also the 0.5mg/ml-1mg/ml polypeptide can be dissolved in the sodium-acetate buffer of pH 4.5, adopt the amido modified method immobilized polypeptide of standard C M5 chip surface, the polypeptide fixed amount is generally 10-50RU when being used for dynamics research, and change with polypeptide molecular weight, generally make its subsequently the protein combination capacity be 500-1000RU.Towards F1 and F2 flow chamber two hours, when baseline stability, with interaction protein in the para-insulin recipient cell signal transduction pathway of variable concentrations simultaneously by F1 and F2, study the interaction of itself and polypeptide, after experiment is finished, with the HBS-EP damping fluid regeneration chip surface that contains 0.5%SDS, if get back to baseline after albumen and the polypeptide effect, regeneration step can save.The polypeptide biochip can be repeatedly used after regeneration.The utility model is built that technology platform can be used for the interactional measurement of other receptor protein and its signal polypeptide and is the polypeptide drugs screening of target spot with this receptor.
Claims (3)
1, a kind of polypeptide biochip structure of simulated receptor signaling molecule, it is characterized in that described polypeptide chip is to go up gilt watch facial mask (2) in glass bottom (1), fixing sephadex layer (3) thereon again, and described sephadex layer (3) is connected with a 6-aminocaprolc acid on the peptide molecule as linking arm (10), the other end of linking arm (10) connects the phosphoric acid gene on the peptide molecule tyrosine residue, the polypeptide of linking arm utilizes the amino fixing means of standard to be fixed in the sephadex layer.
2, by the polypeptide biochip structure of the described simulated receptor signaling molecule of claim 1, it is characterized in that linking arm (10) by standard amino with the sephadex molecular layer on the hydroxyl covalent coupling.
3,, it is characterized in that glass bottom (1), surperficial golden film (2) and fixing sephadex layer are the three-decker of chip surface from the incident light plane to the flow chamber by the polypeptide biochip structure of the described simulated receptor signaling molecule of claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200620048746 CN200989904Y (en) | 2006-12-08 | 2006-12-08 | Polypeptide biochip structure for simulating acceptor signal molecule |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200620048746 CN200989904Y (en) | 2006-12-08 | 2006-12-08 | Polypeptide biochip structure for simulating acceptor signal molecule |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN200989904Y true CN200989904Y (en) | 2007-12-12 |
Family
ID=38940941
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200620048746 Expired - Lifetime CN200989904Y (en) | 2006-12-08 | 2006-12-08 | Polypeptide biochip structure for simulating acceptor signal molecule |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN200989904Y (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009135388A1 (en) * | 2008-05-09 | 2009-11-12 | Zhiping Liu | A molecule detecting system |
| CN104672248A (en) * | 2013-12-03 | 2015-06-03 | 深圳市艾瑞生物科技有限公司 | Covalent coupling agent, chemically modified covalent coupling solid phase carrier containing covalent coupling agent and preparation method and application of carrier |
| CN114200126A (en) * | 2021-12-09 | 2022-03-18 | 牟奕 | Solid phase matrix for detecting N-type penicillin and cephalosporin antibiotic antibodies and preparation method thereof |
-
2006
- 2006-12-08 CN CN 200620048746 patent/CN200989904Y/en not_active Expired - Lifetime
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009135388A1 (en) * | 2008-05-09 | 2009-11-12 | Zhiping Liu | A molecule detecting system |
| CN104672248A (en) * | 2013-12-03 | 2015-06-03 | 深圳市艾瑞生物科技有限公司 | Covalent coupling agent, chemically modified covalent coupling solid phase carrier containing covalent coupling agent and preparation method and application of carrier |
| CN114200126A (en) * | 2021-12-09 | 2022-03-18 | 牟奕 | Solid phase matrix for detecting N-type penicillin and cephalosporin antibiotic antibodies and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Byun | Recent progress and opportunities for nucleic acid aptamers | |
| Heal et al. | Specific interactions between sense and complementary peptides: the basis for the proteomic code | |
| De Crescenzo et al. | Real-time monitoring of the interactions of two-stranded de novo designed coiled-coils: effect of chain length on the kinetic and thermodynamic constants of binding | |
| Jelinek et al. | Carbohydrate biosensors | |
| Schmid et al. | Reversible oriented surface immobilization of functional proteins on oxide surfaces | |
| Linman et al. | Surface plasmon resonance study of protein− carbohydrate interactions using biotinylated sialosides | |
| Sklar et al. | Flow cytometric analysis of ligand-receptor interactions and molecular assemblies | |
| Ziarek et al. | Sulfopeptide probes of the CXCR4/CXCL12 interface reveal oligomer-specific contacts and chemokine allostery | |
| Tse Sum Bui et al. | Molecularly imprinted polymers as synthetic antibodies for protein recognition: The next generation | |
| Durani | Protein design with L-and D-α-amino acid structures as the alphabet | |
| CN200989904Y (en) | Polypeptide biochip structure for simulating acceptor signal molecule | |
| Micksch et al. | Investigation of the peptide adsorption on ZrO2, TiZr, and TiO2 surfaces as a method for surface modification | |
| Hanes et al. | Dual targeting of the chemokine receptors CXCR4 and ACKR3 with novel engineered chemokines | |
| CN101303354A (en) | Biological chips of surface plasma resonating biological sensor, preparation and application | |
| Pesquero et al. | Real-time monitoring and kinetic parameter estimation of the affinity interaction of jArtinM and rArtinM with peroxidase glycoprotein by the electrogravimetric technique | |
| Jørgensen et al. | CCL19 with CCL21-tail displays enhanced glycosaminoglycan binding with retained chemotactic potency in dendritic cells | |
| Li et al. | Chemical synthesis of a full-length G-protein-coupled receptor β2-adrenergic receptor with defined modification patterns at the C-terminus | |
| Oishi et al. | Development of novel CXC chemokine receptor 7 (CXCR7) ligands: selectivity switch from CXCR4 antagonists with a cyclic pentapeptide scaffold | |
| Cho et al. | A vitronectin-derived bioactive peptide improves bone healing capacity of SLA titanium surfaces | |
| Liparoto et al. | Analysis of the role of the interleukin-2 receptor γ chain in ligand binding | |
| CN105954237A (en) | Surface plasma resonance instrument chip and preparation method thereof | |
| CN101506228A (en) | Cysteine-tagged staphylococcal protein G variant | |
| Du et al. | A biomimetic taste biosensor based on bitter receptors synthesized and purified on chip from a cell-free expression system | |
| Fox et al. | Examination of glycosaminoglycan binding sites on the XCL1 dimer | |
| WO2003044530A1 (en) | Sensor system with a reference surface mimicking the detection surface but with low ligand binding capacity |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: CO-PATENTEE TO: HONG KONG CITY UNIV SHENZHEN RESEARCH INSTITUTE ¬ |
|
| CU01 | Correction of utility model patent |
Correction item: Co-patentee Correct: Shenzhen Institute of City University Hong Kong Number: 50 Page: The title page Volume: 23 |
|
| CU03 | Correction of utility model patent gazette |
Correction item: Co-patentee Correct: Shenzhen Institute of City University Hong Kong Number: 50 Volume: 23 |
|
| ERR | Gazette correction |
Free format text: CORRECT: CO-PATENTEE; FROM: NONE ¬ TO: HONG KONG CITY UNIV SHENZHEN RESEARCH INSTITUTE ¬ |
|
| CX01 | Expiry of patent term |
Granted publication date: 20071212 |
|
| EXPY | Termination of patent right or utility model |