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  1. Article ; Online: Sperm cell purification from mock forensic swabs using SOMAmer™ affinity reagents.

    Katilius, Evaldas / Carmel, Andrew B / Koss, Heidi / O'Connell, Dan / Smith, Breanna C / Sanders, Glenn M / LaBerge, Greggory S

    Forensic science international. Genetics

    2018  Volume 35, Page(s) 9–13

    Abstract: We have demonstrated a proof of concept with affinity-based purification of sperm cells from mock forensic samples using SOMAmer™ reagents, DNA-based affinity reagents developed by SomaLogic, Inc. SOMAmer reagents were selected in vitro using whole-cell ... ...

    Abstract We have demonstrated a proof of concept with affinity-based purification of sperm cells from mock forensic samples using SOMAmer™ reagents, DNA-based affinity reagents developed by SomaLogic, Inc. SOMAmer reagents were selected in vitro using whole-cell SELEX to bind specifically with intact, detergent-treated sperm cells. Successful separation of sperm from epithelial cells and their debris was demonstrated using buccal swabs with added semen. Primarily male DNA profiles were generated from sperm cells eluted from the types of cotton swabs typically used for rape kit evidence collection. The quality of sperm DNA isolated from samples purified using SOMAmers is comparable to existing commercially available differential extraction-based methods at higher sperm concentrations. This purification method is simple, offers relatively rapid (<2hr) sperm purification, and can potentially be automated using robotic workstations. This work serves as proof of concept that demonstrates the first use of SOMAmer reagents as affinity ligands to bind sperm cells. With further development, this technique can potentially be used for high-throughput sexual assault forensic casework.
    MeSH term(s) Cell Separation ; DNA/isolation & purification ; DNA Fingerprinting ; Epithelial Cells/cytology ; Female ; Forensic Genetics ; Humans ; Indicators and Reagents ; Male ; Specimen Handling/instrumentation ; Spermatozoa/cytology
    Chemical Substances Indicators and Reagents ; DNA (9007-49-2)
    Language English
    Publishing date 2018-03-27
    Publishing country Netherlands
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2493339-9
    ISSN 1878-0326 ; 1872-4973
    ISSN (online) 1878-0326
    ISSN 1872-4973
    DOI 10.1016/j.fsigen.2018.03.011
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 6764: Show issues Location:
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  2. Article ; Online: Proteomics analysis of cancer exosomes using a novel modified aptamer-based array (SOMAscan™) platform.

    Webber, Jason / Stone, Timothy C / Katilius, Evaldas / Smith, Breanna C / Gordon, Bridget / Mason, Malcolm D / Tabi, Zsuzsanna / Brewis, Ian A / Clayton, Aled

    Molecular & cellular proteomics : MCP

    2014  Volume 13, Issue 4, Page(s) 1050–1064

    Abstract: We have used a novel affinity-based proteomics technology to examine the protein signature of small secreted extracellular vesicles called exosomes. The technology uses a new class of protein binding reagents called SOMAmers® (slow off-rate modified ... ...

    Abstract We have used a novel affinity-based proteomics technology to examine the protein signature of small secreted extracellular vesicles called exosomes. The technology uses a new class of protein binding reagents called SOMAmers® (slow off-rate modified aptamers) and allows the simultaneous precise measurement of over 1000 proteins. Exosomes were highly purified from the Du145 prostate cancer cell line, by pooling selected fractions from a continuous sucrose gradient (within the density range of 1.1 to 1.2 g/ml), and examined under standard conditions or with additional detergent treatment by the SOMAscan™ array (version 3.0). Lysates of Du145 cells were also prepared, and the profiles were compared. Housekeeping proteins such as cyclophilin-A, LDH, and Hsp70 were present in exosomes, and we identified almost 100 proteins that were enriched in exosomes relative to cells. These included proteins of known association with cancer exosomes such as MFG-E8, integrins, and MET, and also those less widely reported as exosomally associated, such as ROR1 and ITIH4. Several proteins with no previously known exosomal association were confirmed as exosomally expressed in experiments using individual SOMAmer® reagents or antibodies in micro-plate assays. Western blotting confirmed the SOMAscan™-identified enrichment of exosomal NOTCH-3, L1CAM, RAC1, and ADAM9. In conclusion, we describe here over 300 proteins of hitherto unknown association with prostate cancer exosomes and suggest that the SOMAmer®-based assay technology is an effective proteomics platform for exosome-associated biomarker discovery in diverse clinical settings.
    MeSH term(s) Biomarkers, Tumor/metabolism ; Cell Line, Tumor ; Exosomes/genetics ; Exosomes/metabolism ; Genes, Essential ; Humans ; Male ; Microarray Analysis/methods ; Nanotechnology ; Prostatic Neoplasms/metabolism ; Proteomics/methods
    Chemical Substances Biomarkers, Tumor
    Language English
    Publishing date 2014-02-06
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2075924-1
    ISSN 1535-9484 ; 1535-9476
    ISSN (online) 1535-9484
    ISSN 1535-9476
    DOI 10.1074/mcp.M113.032136
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 5599: Show issues Location:
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    Jg. 1995 - 2021: Lesesall (2.OG)
    ab Jg. 2022: Lesesaal (EG)

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