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Multiplexed solid-p...
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Darmanis, SpyrosUppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
(författare)
Multiplexed solid-phase proximity ligation assays: Highly specific and parallel protein measurements with DNA sequencing readout
Förlag, utgivningsår, omfång ...
Nummerbeteckningar
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LIBRIS-ID:oai:DiVA.org:uu-145007
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https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-145007URI
Kompletterande språkuppgifter
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Språk:engelska
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Sammanfattning på:engelska
Ingår i deldatabas
Klassifikation
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Ämneskategori:vet swepub-contenttype
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Ämneskategori:ovr swepub-publicationtype
Anmärkningar
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Identification and validation of protein biomarkers is a very important step towards the understanding of the underlying mechanisms of disease, early diagnosis and efficient patient treatment. To carry out this task, methods are needed that would allow us to mine the proteome with sufficient sensitivity and specificity in large sets of samples. We present herein the development of a Multiplexed Proximity Ligation Assay (MultiPLAy), to facilitate efficient protein profiling in a parallel, sensitive and specific manner. We showed that for the simultaneous analysis of 35 proteins MultiPLAy exhibited an improved sensitivity over conventional sandwich assays as well as a smaller susceptibility to background signal increase in the transition from singleplex to multiplex. We used MultiPLAy to identify putative biomarkers in two separate sample cohorts of colorectal cancer (CRC) and cardiovascular disease (CVD) and with the use a novel multivariate analysis approach were able to identify new, as well as already known diagnostic biomarkers. Furthermore we were able to combine MultiPLAy with the use of next-generation sequencing allowing for the first time digital recording of protein profiles in blood. We demonstrated good reproducibility of MultiPLAy coupled to next-generation sequencing, as well as a satisfactory correlation to standard real-time PCR readout. We conclude that MultiPLAy has great potential as a basis for highly multiplexed protein detection assays that can be utilized for the identification of large numbers of proteins or protein variants. This will allow extensive validation of protein expression patterns in biobanked samples and in prospective studies, and can provide a much-needed platform for efficient validation of diagnostic markers for clinical use.
Ämnesord och genrebeteckningar
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multiplex
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proximity ligation assay
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sequencing
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colorectal cancer
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cardiovascular disease
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biomarkers
Biuppslag (personer, institutioner, konferenser, titlar ...)
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Nong, Rachel YuanUppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
(författare)
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Vänelid, JohanUppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
(författare)
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Birgisson, HelgiUppsala universitet,Kolorektalkirurgi
(författare)
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Siegbahn, AgnetaUppsala universitet,Klinisk kemi
(författare)
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Isaksson, MagnusHalo genomics
(författare)
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Ericsson, OlleHalo Genomics
(författare)
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Fredriksson, SimonOlink Biosciences
(författare)
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Glimelius, BengtUppsala universitet,Institutionen för onkologi, radiologi och klinisk immunologi
(författare)
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Wallentin, LarsUppsala universitet,Klinisk kemi
(författare)
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Gustafsson, Mats G.Uppsala universitet,Institutionen för medicinska vetenskaper
(författare)
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Moghaddam, Masood-KamaliUppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
(författare)
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Landegren, UlfUppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
(författare)
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Uppsala universitetInstitutionen för immunologi, genetik och patologi
(creator_code:org_t)
Internetlänk
- Av författaren/redakt...
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Darmanis, Spyros
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Nong, Rachel Yua ...
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Vänelid, Johan
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Birgisson, Helgi
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Siegbahn, Agneta
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Isaksson, Magnus
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visa fler...
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Ericsson, Olle
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Fredriksson, Sim ...
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Glimelius, Bengt
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Wallentin, Lars
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Gustafsson, Mats ...
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Moghaddam, Masoo ...
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Landegren, Ulf
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visa färre...
- Av lärosätet
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Uppsala universitet