Novel Peptide Sequence (“IQ-tag”) with High Affinity for NIR Fluorochromes Allows Protein and Cell Specific Labeling for In Vivo Imaging 英文参考文献.docVIP

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Novel Peptide Sequence (“IQ-tag”) with High Affinity for NIR Fluorochromes Allows Protein and Cell Specific Labeling for In Vivo Imaging 英文参考文献.doc

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Novel Peptide Sequence (“IQ-tag”) with High Affinity for NIR Fluorochromes Allows Protein and Cell Specific Labeling for In Vivo Imaging 英文参考文献

NovelPeptideSequence(‘‘IQ-tag’’)withHighAffinityfor NIRFluorochromesAllowsProteinandCellSpecific LabelingforInVivoImaging KimberlyA.Kelly.,JonathanCarson.,JasonR.McCarthy,RalphWeissleder* CenterforMolecularImagingResearch,MassachusettsGeneralHospital,HarvardMedicalSchool,Boston,Massachusetts,UnitedStatesofAmerica Background. Probes thatallow site-specific protein labeling have become critical tools forvisualizing biological processes. Methods.Hereweusedphagedisplaytoidentifyanovelpeptidesequencewithnanomolaraffinityfornearinfrared(NIR) (benz)indolium fluorochromes. Thedeveloped peptide sequence (‘‘IQ-tag’’) allowsdetection ofNIR dyesin a widerange of assaysincludingELISA,flowcytometry,highthroughputscreens,microscopy,andopticalinvivoimaging.Significance.The described method is expected to have broad utility in numerous applications, namely site-specific protein imaging, target identification,celltracking,anddrugdevelopment. Citation: KellyKA,CarsonJ,McCarthyJR,WeisslederR(2007)NovelPeptideSequence(‘‘IQ-tag’’)withHighAffinityforNIRFluorochromesAllows ProteinandCellSpecificLabelingforInVivoImaging.PLoSONE2(7):e665.doi:10.1371/journal.pone.0000665 INTRODUCTION In the current study we use phage display screening to determine whether peptides with sufficiently high affinity for (benz)indolium-derived fluorochromes could be identified. We found that screening a 7-merlibrary results in convergence onto a single peptide sequence with subnanomolar affinity for these dyes.Furthermore,theutilityofthepeptideforanalyticaltesting, celllabeling,andinvivoimagingisdemonstrated.Thedeveloped peptide sequence should be broadly applicable in a variety of analyticalandimagingapplications(Fig.1). Anumberofadvancesinreporterandimagingtechnologieshave led to the widespread use of fluorescent proteins (FP) and site- specific binders. Fusions offluorescent proteins (e.g. green (GFP) or red fluorescent proteins (RFP)) and target proteins of interest havebeenusedtostudydiversecellularprocess