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Plasma Profiling

National facility

The Plasma Profiling facility offers high throughput analysis of proteins in body fluids using different immunoassay technologies. Protein analysis can be conducted with (i) highly multiplexed antibody bead arrays, (ii) multiplexed kits and technologies from commercial providers, (iii) development of novel ELISA-type assays and (iv) by designing multiplexed assays tailored for user projects. Hosting a unique combination of immunoassay technologies, infrastructure (instrumentation and reagents) and expertise on using the data, the facility works on projects spanning from early discovery towards the implementation of customised immunoassays for validation and translation purposes. Our expertise expands into validation of antibodies, biostatistical data analysis, profiling of interacting proteins as well as consulting the user to fin the most suitable solution for their projects.

 

SERVICES

  • Exploratory protein profiling of body fluids
    • Building customised antibody bead arrays (SBA)
    • Support with study design
    • Support with antibody selection
    • Perform 100 or 384-plex SBA assays (direct sample labelling)
    • Support with data analysis (optional)
  • Development of novel sandwich immunoassays
    • Selection of antibodies and proteins
    • Multiplexed assessment of antibody pairs
    • Development of protocol for body fluid analysis
  • Protein analysis by multiplexed immunoassays
    • Bead based immunoassay (Luminex and commercial providers)
    • Microchannel-based immunoassays (ProteinSimple)
    • Proximity Extension Assays (Olink)
  • Development of immunoassays for protein analysis
  • Consulting on protein analysis using immunoassays

 

Proposed projects are evaluated according to the following model.

1) Initial assessment by facility:

  • Technical feasibility and suitability
  • Capacity and resources requested for the project

2) Project prioritization by committee:

  • Scientific potential
  • Supporting preliminary data
  • Significance of facility specific technique for project
  • Supports facility development (competence and techniques)

APPLICATIONS

The facility offers services for

  • Antibody-based protein analysis.
  • Biomarker discovery in larger sample sets.
  • Multiplexed antibody bead arrays assays.
  • Multiplexed protein analysis based on commercial kits and technologies.
  • Assay development for user defined proteins and projects.
  • Biostatistical processing and analysis of multiplexed data.
  • Expertise in analysis of human body fluids.

EQUIPMENT

  • Reagents
    • 50,000 antibodies from the Human Protein Atlas project, validated on protein arrays
  • Analytical equipment
    • 2x FLEXMAP 3D, Luminex®
    • LX200, Luminex®
    • MAGPIX, Luminex®
    • ELLA, ProteinSimple®,
    • Biomark HD.  from Fluidigm®.
  • Liquid handling
    • FLUENT, Tecan
    • 2x EVO150, Tecan
    • SELMA, CyBio®
  • Magnetic bead handling
    • KingFisher
    • Biotek EL406.

RECENT PROJECTS

  • Profiling 3,000 plasma samples using a customized bead array with 384 HPA antibodies for the discovery of disease-associated proteins. The aims are to identify and validate potential biomarkers; (1) to stratify populations by risk of disease progression, and (2) to monitor organ function in response to treatment.
  • Evaluation and translation of a novel immunoassay for a diabetes-related biomarker.
  • Design of a new immunoassay to study up to 300 proteins for their involvement in protein-protein interactions. We analyzed samples from immunoprecipitations derived from transfected human cells.
  • Development of a new immunoassay to validate proposed biomarker candidates by analyzing lysates of cells collected from 200 cancer patients.

Publications using the facility:

Circulating Carnosine Dipeptidase 1 Associates with Weight Loss and Poor Prognosis in Gastrointestinal Cancer. P Arner et al, PLoS ONE, 2015. 

Heat differentiated complement factor profiling. Hamsten et al, J of Proteomics, 2015.

Affinity proteomics within rare diseases: a BIO-NMD study for blood biomarkers of muscular dystrophies. B Ayoglu et al, EMBO Mol Med, 2014.

Identification of Candidate Serum Proteins for Classifying Well-Differentiated Small Intestinal Neuroendocrine Tumors. S Darmanis et al, PLoS ONE, 2013.