Search: onr:"swepub:oai:DiVA.org:uu-326672" > Analytically Sensit...
Fältnamn | Indikatorer | Metadata |
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000 | 04613naa a2200409 4500 | |
001 | oai:DiVA.org:uu-326672 | |
003 | SwePub | |
008 | 170721s2017 | |||||||||||000 ||eng| | |
024 | 7 | a https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3266722 URI |
024 | 7 | a https://doi.org/10.1373/clinchem.2017.2718332 DOI |
040 | a (SwePub)uu | |
041 | a engb eng | |
042 | 9 SwePub | |
072 | 7 | a ref2 swepub-contenttype |
072 | 7 | a art2 swepub-publicationtype |
100 | 1 | a Ebai, Tongeu Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)toneb991 |
245 | 1 0 | a Analytically Sensitive Protein Detection in Microtiter Plates by Proximity Ligation with Rolling Circle Amplification |
264 | c 2017-09-01 | |
264 | 1 | b Oxford University Press (OUP),c 2017 |
338 | a print2 rdacarrier | |
520 | a BACKGROUND: Detecting proteins at low concentrations in plasma is crucial for early diagnosis. Current techniques in clinical routine, such as sandwich ELISA, provide sensitive protein detection because of a dependence on target recognition by pairs of antibodies, but detection of still lower protein concentrations is often called for. Proximity ligation assay with rolling circle amplification (PLARCA) is a modified proximity ligation assay (PLA) for analytically specific and sensitive protein detection via binding of target proteins by 3 antibodies, and signal amplification via rolling circle amplification (RCA) in microtiter wells, easily adapted to instrumentation in use in hospitals.METHODS: Proteins captured by immobilized antibodies were detected using a pair of oligonucleotide-conjugated antibodies. Upon target recognition, these PLA probes guided oligonucleotide ligation, followed by amplification via RCA of circular DNA strands that formed in the reaction. The RCA products were detected by horseradish peroxidase-labeled oligonucleotides to generate colorimetric reaction products with readout in an absorbance microplate reader.RESULTS: We compared detection of interleukin (IL)-4, IL-6, IL-8, p53, and growth differentiation factor-15 by PLARCA and conventional sandwich ELISA or immuno RCA. PLARCA detected lower concentrations of proteins and exhibited a broader dynamic range compared ELISA and iRCA using the same antibodies. IL-4 and IL-6 were detected in clinical samples at femtomolar concentrations, considerably lower than for ELISA.CONCLUSIONS: PLARCA offers detection of lower protein levels and increased dynamic ranges compared to ELISA. The PLARCA procedure may be adapted to routine instrumentation available in hospitals and research laboratories. | |
650 | 7 | a MEDICIN OCH HÄLSOVETENSKAPx Klinisk medicinx Klinisk laboratoriemedicin0 (SwePub)302232 hsv//swe |
650 | 7 | a MEDICAL AND HEALTH SCIENCESx Clinical Medicinex Clinical Laboratory Medicine0 (SwePub)302232 hsv//eng |
700 | 1 | a de Oliveira, Felipe Marques Souzau Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)felma546 |
700 | 1 | a Löf, Lizau Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)lizlo931 |
700 | 1 | a Wik, Lottau Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)lowik226 |
700 | 1 | a Schweiger, Carolineu Charité Comprehensive Cancer Center, University of Berlin, Berlin, Germany; Institute of Pathology, Medical University of Graz, Graz, Austria;4 aut |
700 | 1 | a Larsson, Andersu Uppsala universitet,Biokemisk struktur och funktion4 aut0 (Swepub:uu)andlarss |
700 | 1 | a Keilholtz, Ulrichu Institute of Pathology, Medical University of Graz, Graz, Austria;4 aut |
700 | 1 | a Haybaeck, Johannesu Charité Comprehensive Cancer Center, University of Berlin, Berlin, Germany; Department of Pathology, Otto von Guericke University Magdeburg, Magdeburg, Germany.4 aut |
700 | 1 | a Landegren, Ulfu Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)ulfland |
700 | 1 | a Kamali-Moghaddam, Masoodu Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)masookam |
710 | 2 | a Uppsala universitetb Science for Life Laboratory, SciLifeLab4 org |
773 | 0 | t Clinical Chemistryd : Oxford University Press (OUP)g 63:9, s. 1497-1505q 63:9<1497-1505x 0009-9147x 1530-8561 |
856 | 4 | u https://academic.oup.com/clinchem/article-pdf/63/9/1497/32647318/clinchem1497.pdf |
856 | 4 8 | u https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-326672 |
856 | 4 8 | u https://doi.org/10.1373/clinchem.2017.271833 |
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