CN1142610A - Method for analyzing non-covalent effect between biological composition - Google Patents
Method for analyzing non-covalent effect between biological composition Download PDFInfo
- Publication number
- CN1142610A CN1142610A CN 95107971 CN95107971A CN1142610A CN 1142610 A CN1142610 A CN 1142610A CN 95107971 CN95107971 CN 95107971 CN 95107971 A CN95107971 A CN 95107971A CN 1142610 A CN1142610 A CN 1142610A
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- CN
- China
- Prior art keywords
- electrophoresis
- biological
- radioactive
- biotin
- mark
- 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.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 17
- 230000000694 effects Effects 0.000 title claims description 5
- 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 22
- 239000007787 solid Substances 0.000 claims abstract description 12
- 229960002685 biotin Drugs 0.000 claims abstract description 11
- 235000020958 biotin Nutrition 0.000 claims abstract description 11
- 239000011616 biotin Substances 0.000 claims abstract description 11
- 230000002285 radioactive effect Effects 0.000 claims abstract description 9
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 6
- 238000001962 electrophoresis Methods 0.000 claims abstract description 6
- 108090001008 Avidin Proteins 0.000 claims abstract description 3
- 108010090804 Streptavidin Proteins 0.000 claims abstract description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 6
- 230000003993 interaction Effects 0.000 claims description 6
- 230000001745 anti-biotin effect Effects 0.000 claims description 4
- 238000001502 gel electrophoresis Methods 0.000 claims description 4
- 230000006287 biotinylation Effects 0.000 claims description 3
- 238000007413 biotinylation Methods 0.000 claims description 3
- 238000004132 cross linking Methods 0.000 claims description 3
- 239000007791 liquid phase Substances 0.000 claims description 3
- 238000004458 analytical method Methods 0.000 claims description 2
- 239000012472 biological sample Substances 0.000 claims description 2
- 238000013016 damping Methods 0.000 claims description 2
- 239000012530 fluid Substances 0.000 claims description 2
- 238000011534 incubation Methods 0.000 claims description 2
- 239000000523 sample Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 abstract 2
- 239000000126 substance Substances 0.000 abstract 2
- 230000003139 buffering effect Effects 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- 238000001514 detection method Methods 0.000 description 2
- 241000094396 Bolitoglossa carri Species 0.000 description 1
- 241001597008 Nomeidae Species 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000009878 intermolecular interaction Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
A method for analyzing noncovalent action between biologic components includes such steps as use of biotin reagent and radioactive isotop to label different components, putting labeled components in liquid for reacting on each other, adding avidin or like streptavidin covalently crosslinked to solid supporter, mixing and waiting for a certain time, washing solid substance with proper buffering liquid, collecting solid substance, preparing gel sample for electrophoresis, doing electrophoresis, radioactive self-developing of gel after electrophoresis, and comparation of self-developed results to determine which isotop labeled moleculae have special mutual action on biotin labeled ones.
Description
What the present invention relates to is a kind of method of analyzing non-covalent effect between biological composition, particularly unites use, the different biological components of mark with biotin and radioactive isotope, to study the method for biological intermolecular interaction.
In the biological study field, we often need to determine will produce which type of interaction between the biological components of some characteristic the unknown.Known research method is still undesirable aspect accuracy.
Known in the test organisms molecular engineering people usually adopt a kind of labelled with radioisotope method that certain biological components is carried out spike to detect.The technology of another test organisms molecule is the derivant mark biomolecule with biotin (biotin), the detection technique that biological components to be detected is separated.
The objective of the invention is radioisotope method in the detection of biological molecular engineering and biotin method are united use in the analyzing non-covalent effect between biological composition checkout procedure, thereby a kind of method that can fully not interact and analyze between two groups of biological components of understanding as yet its biological property exactly is provided.
The method that the present invention proposes is made of following steps:
1, with biotinylation reagent and the different biological components of radioactive isotope difference mark;
2, these the two kinds biological components that mark is good are placed liquid phase, it is interacted under certain conditions;
3, add antibiotin (avidin) or other analog streptavidin of covalent cross-linking then, mix the back incubation after the regular hour, fully wash solid formation with suitable damping fluid again in solid support;
4, collect solid formation, preparative gel electrophoresis sample, thereby make gel electrophoresis;
5, do radioactive automatic developing with the gel behind the above-mentioned electrophoresis;
6, the result to the autography of the biological sample handled through above-mentioned steps compares analysis, with the molecule of definite those labelled with radioisotope with there being specific interaction between the biotin labeled molecule.
Specifically, be that B. carries out mark with biotin chemistry reagent on component A if one of them biological components is A, another biological components, and on B component, carry out mark with radioactive isotope.In liquid phase, make the two-way interaction then.If the two can interact, then will produce a product A+B.In this product, promptly contain biotin reagent and also contain isotope reagent.So when adding covalent cross-linking in the antibiotin of solid support, above-mentioned product then will generate solid formation with the antibiotin effect because of it contains biotin, and this solid formation is carried out can producing gel behind the electrophoresis.Owing to this gel is derived by A+B, thereby wherein also contain isotope.Whether therefore can detect it at an easy rate with radioactive automatic developing exists.Can analyze A and B thus and whether produce noncovalent interaction.
By above-mentioned narration as can be seen, adopt this method can utilize the advantage of the easily separated and easy purifying of the biomolecule of good advantage of the high sensitivity of labelled with radioisotope method and specificity and biotinylation mark simultaneously, thereby the method for interaction property between a kind of component of test organisms exactly is provided.
Claims (1)
1, a kind of method of analyzing non-covalent effect between biological composition is characterized in that this method comprises the steps:
1., with biotinylation reagent and the different biological components of radioactive isotope difference mark;
2., these the two kinds biological components that mark is good are placed liquid phase, it is interacted under certain conditions;
3., then add antibiotin (avidin) or other analog streptavidin of covalent cross-linking, mix the back incubation after the regular hour, fully wash solid formation with suitable damping fluid again in solid support;
4., collect solid formation, preparative gel electrophoresis sample, thereby make gel electrophoresis;
5., do radioactive automatic developing with the gel behind the above-mentioned electrophoresis;
6., the result of the autography of the biological sample handled through above-mentioned steps is compared analysis, with the molecule of definite those labelled with radioisotope with there being specific interaction between the biotin labeled molecule.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 95107971 CN1142610A (en) | 1995-08-08 | 1995-08-08 | Method for analyzing non-covalent effect between biological composition |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 95107971 CN1142610A (en) | 1995-08-08 | 1995-08-08 | Method for analyzing non-covalent effect between biological composition |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1142610A true CN1142610A (en) | 1997-02-12 |
Family
ID=5076556
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 95107971 Pending CN1142610A (en) | 1995-08-08 | 1995-08-08 | Method for analyzing non-covalent effect between biological composition |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1142610A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1052510A4 (en) * | 1998-11-06 | 2002-09-25 | Iatron Lab | NOVEL COMPULS CONTAINING CROSSLINKED AVIDIN, ANALYTICAL PROCESS IN WHICH CROSSLINKED AVIDINE IS USED, REAGENTS AND ANALYSIS KITS |
-
1995
- 1995-08-08 CN CN 95107971 patent/CN1142610A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1052510A4 (en) * | 1998-11-06 | 2002-09-25 | Iatron Lab | NOVEL COMPULS CONTAINING CROSSLINKED AVIDIN, ANALYTICAL PROCESS IN WHICH CROSSLINKED AVIDINE IS USED, REAGENTS AND ANALYSIS KITS |
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| C06 | Publication | ||
| PB01 | Publication | ||
| C01 | Deemed withdrawal of patent application (patent law 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |