Sökning: onr:"swepub:oai:DiVA.org:uu-145007" >
Multiplexed solid-p...
Multiplexed solid-phase proximity ligation assays: Highly specific and parallel protein measurements with DNA sequencing readout
-
- Darmanis, Spyros (författare)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
-
- Nong, Rachel Yuan (författare)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
-
- Vänelid, Johan (författare)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
-
visa fler...
-
- Birgisson, Helgi (författare)
- Uppsala universitet,Kolorektalkirurgi
-
- Siegbahn, Agneta (författare)
- Uppsala universitet,Klinisk kemi
-
- Isaksson, Magnus (författare)
- Halo genomics
-
- Ericsson, Olle (författare)
- Halo Genomics
-
- Fredriksson, Simon (författare)
- Olink Biosciences
-
- Glimelius, Bengt (författare)
- Uppsala universitet,Institutionen för onkologi, radiologi och klinisk immunologi
-
- Wallentin, Lars (författare)
- Uppsala universitet,Klinisk kemi
-
- Gustafsson, Mats G. (författare)
- Uppsala universitet,Institutionen för medicinska vetenskaper
-
- Moghaddam, Masood-Kamali (författare)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
-
- Landegren, Ulf (författare)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi,Molecular medicine
-
visa färre...
-
(creator_code:org_t)
- Engelska.
- Relaterad länk:
-
https://urn.kb.se/re...
Abstract
Ämnesord
Stäng
- 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.
Nyckelord
- multiplex
- proximity ligation assay
- sequencing
- colorectal cancer
- cardiovascular disease
- biomarkers
Publikations- och innehållstyp
- vet (ämneskategori)
- ovr (ämneskategori)
- Av författaren/redakt...
-
Darmanis, Spyros
-
Nong, Rachel Yua ...
-
Vänelid, Johan
-
Birgisson, Helgi
-
Siegbahn, Agneta
-
Isaksson, Magnus
-
visa fler...
-
Ericsson, Olle
-
Fredriksson, Sim ...
-
Glimelius, Bengt
-
Wallentin, Lars
-
Gustafsson, Mats ...
-
Moghaddam, Masoo ...
-
Landegren, Ulf
-
visa färre...
- Av lärosätet
-
Uppsala universitet