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Types of Protein Microarray and Antibody Chips Typer af Protein microarray og Antibody Chips

Protein and Antibody Microarrays Protein og Antibody Microarrays

Protein Chips and Antibody Microarrays - Glass Slides, Microwell/Nanowell Protein-chips og Antibody Microarrays - glasslides, Microwell / Nanowell

Two major protein chip formats are now used, including glass slides and nanowells (8,27).  Due to the large amounts of slide adherence methods only the most common and major types will be mentioned here. To vigtige protein chip formater er nu anvendes, herunder glasslides og nanowells (8,27). På grund af de store mængder af dias tilslutning metoder kun de mest almindelige og store typer vil blive nævnt her.
It is important that protein chips retain proteins in an active state at high densities, are compatible with most commercial arrayers and scanners, and can be printed in such a fashion that the proteins remain in a moisturized environment (8,27). Det er vigtigt, at protein chips beholde proteiner i en aktiv tilstand ved høje tætheder, er forenelige med de fleste kommercielle arrayers og scannere, og kan trykkes på en sådan måde, at de proteiner forbliver i en fugtes miljøet (8,27). (See Figure 2). (Se figur 2).

Glass Slide Chips Glas Slide Chips
Glass slides have the advantage that they are compatible with standard microarray equipment and detection equipment used for DNA chips.  They are also inexpensive.  The majority of studies are now using glass slides.  However, they have a high evaporation rate and are susceptible to possible cross-contamination (8,27). Glasslides har den fordel, at de er forenelige med standard microarray udstyr og afsløring udstyr, der bruges til DNA-chips. De er også billige. De fleste undersøgelser er nu brug af glas lysbilleder. Men de har en høj fordampning sats og er modtagelige for muligt på tværs - forurening (8,27).
The first strategies for the creation of protein arrays on glass were developed by Mirzabekov et al..  Arrays were produced by immobilizing proteins in tiny gel pockets that were attached to the glass surface.  A variety of immunoassays, antigen detection, and enzymatic assays were carried out.  Due to the 3D matrix structure, protein immobilization was very efficient (28-30). Den første strategier for oprettelse af protein RAID-glas blev udviklet af Mirzabekov et al .. Arrays blev produceret af immobilizing proteiner i bittesmå gel lommer, der var fastgjort til glas overflade. Forskellige immunassays, antigen afsløring og enzymatiske assays blev foretaget ud. På grund af 3D-matrix-struktur, protein immobilisering var meget effektivt (28-30).
In another study, proteins were attached to a glass surface activated with a crosslinking agent that reacts with primary amines (31).  Proteins were spotted in a 40% glycerol solution which keeps proteins in a wet environment and prevents dehydration.  To determine that their protein microarrays were feasible for biochemical assays, they tested three known protein-protein interactions, three known kinase-substrate reactions, and three known protein-ligand interactions using fluorophore-tagged proteins, radiolabeled ATP, and synthetic ligands coupled to fluorescently labeled bovine serum albumin (BSA), respectively.  In most of the experiments, small numbers of proteins were spotted however in one experiment they identified a single spot within an array of 10,000 spots containing one other protein.  This work demonstrated the potential of protein microarrays in large-scale biochemical assays however their study analyzed a very small number of proteins and novel activities were not identified. I en anden undersøgelse, proteiner var knyttet til et glas overflade aktiveres med et crosslinking agent, der reagerer med primære aminer (31). Proteiner blev opdaget i en 40% glycerol løsning, som holder proteiner i et vådt miljø og forhindrer dehydrering. At fastslå, at deres protein microarrays var muligt for biokemiske assays, de testede tre kendte protein-protein interaktioner, tre kendte kinase-substrat reaktioner, og tre kendte protein-ligand interaktioner bruger fluorophore-mærkede proteiner, radioaktivt ATP, og syntetisk ligands koblet til fluorescently stemplet bovint serum albumin ( BSA), respectively. I de fleste af de eksperimenter, små numre af proteiner var plettede dog i et eksperiment, de har udpeget en enkelt stedet inden for en vifte af 10000 spots indeholdende en anden protein. Dette arbejde viste potentiale af protein microarrays i stor-skala biokemiske assays dog deres undersøgelse analyseres en meget lille antal af proteiner og nye aktiviteter blev ikke identificeret.
Another study used glass slides for the detection of several protein antigens.  Analytes were spotted onto a treated glass surface at high density using a hand-spotting device or a microarray robot.  The spotted analytes were detected using antibodies attached to an oligonucleotide primer and a rolling circle amplification reaction.  The technique had a high sensitivity, a wide dynamic range, and excellent spot-to-spot reproducibility En anden undersøgelse anvendes glasslides til påvisning af flere protein-antigener. Analysander blev spottet på en behandlet glas overflade med høj massefylde ved hjælp af en hånd-spotting anordning eller et microarray robot. Plettede analysander blev opdaget ved hjælp af antistoffer knyttet til en oligonukleotid primer og et rullende cirkel amplifikation reaktion. Teknikken havde en høj følsomhed, et bredt dynamikområde og fremragende spot-til-spot reproducerbarhed
(32).
Most groups now directly array proteins and antibodies onto plain glass slides (31,33-35) and spotting is carried out in a humidity-controlled environment (8). De fleste grupper nu direkte array proteiner og antistoffer på almindeligt glasslides (31,33-35) og genkendelse foregår i en luftfugtighed-kontrolleret miljø (8).

Microwell/Nanowell Chips Microwell / Nanowell Chips
Compatible with standard microarray and detection equipment however alignment is required.  This method is versatible for solution-based assays and multiple-component reactions.  Evaporation is reduced and there is no cross-contamination.  Also these chips are relatively inexpensive (8,27). Kompatibel med standard microarray og opsporingsudstyr dog tilpasning er nødvendig. Denne metode er versatible til opløsning-baserede assays og multiple-komponent reaktioner. Fordampningen er reduceret, og der er ingen krydskontaminering. Også disse chips er relativt billigt (8,27).
Zhu et al., fabricated an open structure, namely nanowells on a polydimethylsiloxane (PDMS) surface supported by the standard glass slides (36). Zhu et al., Opdigtede en åben struktur, nemlig nanowells på en Polydimethylsiloxan (PDMS) overflade understøttes af standardkernen glasslides (36).
These chips consist of an array of microwells in a disposable silicone elastomer, poly(dimethylsiloxane) (PDMS) (37). Disse chips bestå af en vifte af microwells i en engangsemballage silikone elastomer, poly (dimethylsiloxane) (PDMS) (37). Microwell arrays allow small volumes of different analytes to be densely packed on a single chip, yet remain physically segregated during subsequent batch processing. Microwell arrays tillade små mængder af forskellige analysander at være tæt pakket på en enkelt chip, men alligevel holdes fysisk adskilte under den efterfølgende serie forarbejdning. Proteins were covalently attached to the wells using a crosslinker 3-glycidoxypropyltrimethoxysilane (GPTS) (38). Proteiner blev covalently knyttet til brøndene ved hjælp af en crosslinker 3-glycidoxypropyltrimethoxysilane (GPTS) (38).
Captured molecules can be easily recovered from the nanowells.  When covered with gold in the nanowells, it is expected that high-throughput mass spectrometry and surface plasmon resonance analyses can be performed.  The greatest disadvantage of this technique is that specialzed equipment is required to load the nanowells at high density (8,27). Captured molekyler, der let kan genvindes fra nanowells. Når beklædt med guld i nanowells forventes det, at Højkapacitetsforskning mass spectrometry og overflade plasmon resonans-analyser kan udføres. Den største ulempe ved denne teknik er, at specialzed udstyr er påkrævet for at indlæse Det nanowells med høj massefylde (8,27).

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Introduction and Background to Protein Chips and Antibody Chips. Indledning og baggrund for protein-chips og Antibody Chips.

References for Protein and Antibody Microarrays Referencer for protein og Antibody Microarrays