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| - | ~~NOTOC~~ | ||
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| - | **Proteomics Area** | ||
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| - | ====== Surface Plasmon Resonance (SPR - Biacore) ====== | ||
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| - | <WRAP tabs> | ||
| - | * [[en:aree:proteomica:biacore:start|Abstract]] | ||
| - | * [[en:aree:proteomica:biacore:metodica|Methodology]] | ||
| - | * [[en:aree:proteomica:biacore:applicazioni|Applications]] | ||
| - | </WRAP> | ||
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| - | ==== Applications ==== | ||
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| - | Typical applications are: | ||
| - | * affinity and kinetics; | ||
| - | * quantitative titration of analyte; | ||
| - | * specificity; | ||
| - | * compliance to a Reference Standard; | ||
| - | * termodynamics. | ||
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| - | {{ :aree:proteomica:biacore:ambiti_di_misura.png?600 |}} | ||
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| - | ===== Example: kinetic test ===== | ||
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| - | Here is presented an example of consecutive injections analysis. As presented above the instrument can perform sequential analysis of the kinetic. Here we stratified a specific epitope to the sensor chip and then we tested the affinity for its specific monoclonal antibody. | ||
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| - | {{ :aree:proteomica:biacore:test_cinetici.jpeg?600 |}} | ||
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| - | ===== Example: thermodinamic test ===== | ||
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| - | {{ :aree:proteomica:biacore:test_termodinamici.png?600 |}} | ||
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| - | ===== Sensor chip from the manufacturer ===== | ||
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| - | ^ Sensor chip type ^ Characteristics ^ Examples ^ | ||
| - | | General purpose | Sensor chips with carboxyl or carboxymethyl groups on the chip surface for ligand attachment. | Sensor Chips CM3, CM4, CM5, CM7 and C1 | | ||
| - | | Tagged ligand capture | Pre-immobilized with capturing molecules specific for tagged ligands | Sensor Chips SA and NTA \\ NTA Reagent Kit, Biotin CAPture Kit, GST Capture Kit | | ||
| - | | Antibody capture | Pre-immobilized with capturing molecules specific for antibodies. | Sensor Chips Protein A, Protein G, Protein L \\ Human Antibody Capture Kit, \\ Mouse antibody capture Kit, \\ Human Fab Capture Kit. | | ||
| - | | Lipid attachment | Hydrophobic surface for attachment of lipid bilayers or liposomes. | Sensor Chips HPA, L1 | | ||
| - | | Customizable | Unmodified gold surface for preparation of customized surfaces((Customization of unmodified sensor chip surfaces is the responsibility of the user. GE does not provide customization services or support for customized surfaces.)). | Sensor Chip Au \\ SIA Kit | | ||
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| - | As reported earlier the sensor chips are a pivotal element in the experiment design and have to be well chosen to succeed. | ||
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| - | Oltre ai già citati sensori della serie CM- ne esistono di altri più adeguati a legare i lipidi e le molecole idrofobiche quali ad esempio micelle e liposomi. Sono appunto i sensori HPA per monolayers lipidici e quelli di tipo L1 con ancore molecolari idrofobiche che consentono di immobilizzare doppi strati lipidici, frammenti di membrane plasmatiche, ecc. | ||
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| - | Over the yet described CM chip sensors, there are many others more adequate in lipidomic studies for example in the HPA-series a flat hydrophobic surface allow to obtain lipid monolayer, in other cases, it is possible to use L1-series having molecular anchors able to immobilize bilayers or liposomes, plasmic membrane fragments etc. | ||
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| - | Finally, other more specific chips are pre-casted by specific biomolecules such as antibodies, Protein A, G, L streptavidin or Fc from specific immunoglobulins. | ||
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