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Applications of Protein Microarrays Anvendelser af Protein Microarrays

Protein and Antibody Microarrays Protein og Antibody Microarrays

Protein Chip Applications Protein Chip Applications

Applications of Protein Chips Anvendelser af Protein Chips
1) Proteomics 1) Proteomics
Protein chip technologies will provide a powerful, high-throughput and versatile tool for the genome-scale analysis of gene function (see Figure 4).  Enzyme activity, protein–protein and protein–nucleic-acid interactions, and small-molecule drug interactions may all be analyzed directly on the protein level (77,78).  Arrays may be engineered to address protein identification, quantitation, and affinity studies.  A profiling array may quantitate levels of specific proteins on a global scale allowing for a comparison of normal and disease states.  An affinity array may analyze the interactions of peptides, proteins, oligonucleotides, sugars, lipids, or small molecules and chemicals with immobilized proteins such as receptors, enzymes, or antibodies (8). Protein chip teknologier vil give en kraftfuld og høj overførselshastighed og alsidigt værktøj for genom-skala analyse af gen-funktion (se figur 4). Enzyme aktivitet, protein-protein og protein-nukleinsyre-syre interaktioner, og små-molekyle lægemiddelinteraktioner mai alle skal analyseres direkte på protein-niveau (77,78). Arrays kan blive manipuleret til at behandle protein identifikation, quantitation, og affinitet undersøgelser. En profilering array mai kvantificere niveauer af specifikke proteiner på globalt plan giver mulighed for en sammenligning af den normale og sygdom stater. en affinitet array mai analysere samspillet af peptider, proteiner, oligonucleotides, sukker, fedt eller små molekyler og kemikalier med ubevaegeligt proteiner såsom receptorer, enzymer eller antistoffer (8).
Currently, the rate-limiting step is the production of large numbers of proteins. I øjeblikket er det hastighedsbegrænsende trin er produktionen af et stort antal proteiner. The ability to automate protein production and proteins fused to high-affinity tags will greatly expedite protein-chip development. Evnen til at automatisere protein produktion og proteiner smeltet til høj affinitet tags i høj grad vil fremskynde protein-chip-udvikling.  High-density chips containing large sets of proteins or even entire proteomes will allow the high-throughput analysis of biochemical activities, protein–protein interactions and post-translational modifications, such as phosphorylation, dephosphorylation, protein methylation, and ubiquitination. High-density chips indeholder store sæt af proteiner eller endda hele proteomes vil gøre det muligt for high-throughput analyse af biokemiske aktiviteter, protein-protein interaktioner og post-translationel ændringer, såsom fosforylering, dephosphorylation, proteiner methylering, og ubiquitination.

The ultimate goal of proteomics is to the study biochemical activities of every protein encoded by an organism or proteome.  A landmark study conducted prepared the first proteome chip by cloning ~94% (>5800 of 6200) of the yeast open reading frames in a yeast expression vector which expressed the proteins as N-terminal GST-His x6 double tagged fusions.  A high-throughput yeast protein purification method was developed to individually purify proteins. Det ultimative mål for proteomik er til undersøgelsen biokemiske aktiviteter af enhver protein kodet ved en organisme eller proteomanalyse. En skelsættende undersøgelse udarbejdet den første proteomanalyse chip ved kloning ~ 94% (> 5800 i 6200) af gær åben læsning rammer i en gær udtryk vektoren, der udtrykte proteiner som N-terminal gst-Hans x6 dobbelt mærkede fusions. En høj overførselshastighed gær protein rensning metode blev udviklet til individuelt at rense proteiner. 80% of yeast proteins were full length and of sufficient quantity to be detectable by most assay types. 80% af gær proteiner var fulde længde og i tilstrækkelig mængde til at kunne spores i de fleste assay typer. The proteins were then purified using the GST tags and were then attached to Ni-NTA-coated glass slides using the HisX6 tags.  In addition to identifying known interactions, 33 novel binding proteins were detected.  150 novel lipid-binding proteins were also identified.  This study demonstrated that an entire proteome can be immobilized on a glass surface to directly screen for interactions with proteins and small molecules (26). Proteinerne blev derefter renset ved hjælp af GST tags og blev derefter knyttet til Ni-NTA-coatede glasslides bruge HisX6 tags. Ud over at identificere kendte interaktioner, 33 nye bindende proteiner blev opdaget. 150 nye lipid-bindende proteiner blev også identificeret. Denne undersøgelse viste, at en hel proteomanalyse kan være immobiliseret på en glasplade til direkte at skærmen for interaktioner med proteiner og små molekyler (26).
The coupling of mass-spectrometry and protein chips will have wide applications in identifying players in protein–protein interactions, and also in drug discovery (80). Den sammenkobling af masse-spektrometri og protein-chips vil have bred ansøgninger med at identificere aktørerne i protein-protein interaktioner, og også i lægemiddeludvikling (80).  Proteins and small-molecule ligands bound to proteins immobilized on chip can be identified using matrix-assisted laser desorption/ionisation time of flight (MADLI-TOF) mass spectroscopy. Proteiner og små-molekyle ligands bundet til proteiner immobiliseret på chippen kan identificeres ved hjælp af matrix-assisteret laser desorption / ionisation tidspunktet for flyvning (MADLI-TOF) masse spektroskopi.  Microwell formats are particularly suited for this purpose. Microwell formater er særligt velegnede til dette formål. Thus, molecules and proteins that specifically bind to many different proteins can be identified and this information can be used to deduce molecular networks and pathways. Således molekyler og proteiner, som specifikt binder sig til mange forskellige proteiner kan identificeres, og disse oplysninger kan bruges til at udlede molekylære netværk og ophold.
One area that will require technological improvements is the analysis of membrane proteins. Et område, som vil kræve teknologiske forbedringer er analysen af membran proteiner.  A large amount of proteins are likely to be membrane-bound, since as many as one third of all yeast proteins are membrane proteins or secreted proteins (81). En stor mængde af proteiner kan forventes at være membran-bundet, da så mange som en tredjedel af alle gær proteiner er membran proteiner eller udskilles proteiner (81). Due to the fact that many of these proteins are active when in membranes, it therefore may be necessary to purify or reconstitute them with associated lipids. På grund af det faktum, at mange af disse proteiner er aktive, når de er i membraner, er det derfor kan være nødvendigt at rense eller rekonstruere dem med tilhørende lipider. However, this may not be so difficult. Men dette kan ikke være så svært. One group was able to immobilize biotinylated membranes that contain the G-protein-coupled receptor rhodopsin on a gold-coated glass surface, and establish a functional assay for that protein (82). En gruppe var i stand til at immobilisere biotinylerede membraner, der indeholder G-protein-koblede receptor Rhodopsin på en guld-coatede glas overflade, og etablere et funktionelt assay for, at protein (82).  Similar procedures may make it possible to analyze membrane proteins in a chip format. Lignende procedurer kan gøre det muligt at analysere membran proteiner i en chip-format.
2) Diagnostics 2) Diagnostics
Another area which will benefit from protein areas is diagnostics. Et andet område, der vil drage fordel af protein områder er diagnostik. Highly parallel analysis on arrays will allow determination of disease markers (eg tumour markers) in extracts with only a minimum of biopsy (sample) material, creating new possibilities for monitoring disease (cancer) treatment and therapy (83). Stærkt parallel analyse på arrays vil tillade bestemmelse af sygdommen markører (f.eks tumor markører) i uddrag med kun et minimum af biopsi (prøve) materiale, hvilket skaber nye muligheder for overvågning af sygdommen (kræft) behandling og terapi (83).

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

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

Types of Antibody and Protein Chips Typer af antistof og Protein Chips