| 1. |
- Darmanis, Spyros, et al.
(författare)
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ProteinSeq : high-performance proteomic analyses by proximity ligation and next generation sequencing
- 2011
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 6:9, s. e25583-
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Tidskriftsartikel (refereegranskat)abstract
- Despite intense interest, methods that provide enhanced sensitivity and specificity in parallel measurements of candidate protein biomarkers in numerous samples have been lacking. We present herein a multiplex proximity ligation assay with readout via realtime PCR or DNA sequencing (ProteinSeq). We demonstrate improved sensitivity over conventional sandwich assays for simultaneous analysis of sets of 35 proteins in 5 μl of blood plasma. Importantly, we observe a minimal tendency to increased background with multiplexing, compared to a sandwich assay, suggesting that higher levels of multiplexing are possible. We used ProteinSeq to analyze proteins in plasma samples from cardiovascular disease (CVD) patient cohorts and matched controls. Three proteins, namely P-selectin, Cystatin-B and Kallikrein-6, were identified as putative diagnostic biomarkers for CVD. The latter two have not been previously reported in the literature and their potential roles must be validated in larger patient cohorts. We conclude that ProteinSeq is promising for screening large numbers of proteins and samples while the technology can provide a much-needed platform for validation of diagnostic markers in biobank samples and in clinical use.
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| 2. |
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| 4. |
- Ke, Rongqin, et al.
(författare)
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Improving Precision of Proximity Ligation Assay by Amplified Single Molecule Detection
- 2013
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 8:7
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Tidskriftsartikel (refereegranskat)abstract
- Proximity ligation assay (PLA) has been proven to be a robust protein detection method. The technique is characterized by high sensitivity and specificity, but the assay precision is probably limited by the PCR readout. To investigate this potential limitation and to improve precision, we developed a digital proximity ligation assay for protein measurement in fluids based on amplified single molecule detection. The assay showed significant improvements in precision, and thereby also detection sensitivity, over the conventional real-time PCR readout.
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| 5. |
- Lundberg, Martin, et al.
(författare)
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Multiplexed Homogeneous Proximity Ligation Assays for High-throughput Protein Biomarker Research in Serological Material
- 2011
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Ingår i: Molecular & Cellular Proteomics. - 1535-9476 .- 1535-9484. ; 10:4, s. M110.004978-
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Tidskriftsartikel (refereegranskat)abstract
- A high throughput protein biomarker discovery tool has been developed based on multiplexed proximity ligation assays in a homogeneous format in the sense of no washing steps. The platform consists of four 24-plex panels profiling 74 putative biomarkers with sub-pM sensitivity each consuming only 1 mu l of human plasma sample. The system uses either matched monoclonal antibody pairs or the more readily available single batches of affinity purified polyclonal antibodies to generate the target specific reagents by covalently linking with unique nucleic acid sequences. These paired sequences are united by DNA ligation upon simultaneous target binding forming a PCR amplicon. Multiplex proximity ligation assays thereby converts multiple target analytes into real-time PCR amplicons that are individually quantified using microfluidic high capacity qPCR in nano liter volumes. The assay shows excellent specificity, even in multiplex, by its dual recognition feature, its proximity requirement, and most importantly by using unique sequence specific reporter fragments on both antibody-based probes. To illustrate the potential of this protein detection technology, a pilot biomarker research project was performed using biobanked plasma samples for the detection of colorectal cancer using a multivariate signature.
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| 6. |
- Strandroth, Johan, 1978, et al.
(författare)
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Correlation between Euro NCAP Pedestrian Test Results and Injury Severity in Injury Crashes with Pedestrians and Bicyclists in Sweden
- 2014
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Ingår i: SAE Technical Papers. - 400 Commonwealth Drive, Warrendale, PA, United States : SAE International. - 0148-7191 .- 2688-3627. ; 2014-November:November
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Tidskriftsartikel (refereegranskat)abstract
- Pedestrians and bicyclists account for a significant share of deaths and serious injuries in the road transport system. The protection of pedestrians in car-to-pedestrian crashes has therefore been addressed by friendlier car fronts and since 1997, the European New Car Assessment Program (Euro NCAP) has assessed the level of protection for most car models available in Europe. In the current study, Euro NCAP pedestrian scoring was compared with real-life injury outcomes in car-to-pedestrian and car-to-bicyclist crashes occurring in Sweden. Approximately 1200 injured pedestrians and 2000 injured bicyclists were included in the study. Groups of cars with low, medium and high pedestrian scores were compared with respect to pedestrian injury severity on the Maximum Abbreviated Injury Scale (MAIS)-level and risk of permanent medical impairment (RPMI). Significant injury reductions to both pedestrians and bicyclists were found between low and high performing cars. For pedestrians, the reduction of MAIS2+, MAIS3+, RPMI1+ and RPMI10+ ranged from 20-56% and was significant on all levels except for MAIS3+ injuries. Pedestrian head injuries had the highest reduction, 80-90% depending on level of medical impairment. For bicyclist, an injury reduction was only observed between medium and high performing cars. Significant injury reductions were found for all body regions. It was also found that cars fitted with autonomous emergency braking including pedestrian detection might have a 60-70% lower crash involvement than expected. Based on these results, it was recommended that pedestrian protection are implemented on a global scale to provide protection for vulnerable road users worldwide.
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