Plasma Profiling

National facility

Building on the unique resource of > 48,000 polyclonal antibodies, the Plasma profiling facility offers high throughput analysis of proteins in body fluids. With protein targets proposed by the user, up to 380 antibodies can be combined in one array to analyze 384 or more samples in one experiment, using only µl of sample. The facility also offers tailored immunoassays for validation of protein capture, profiling of potential interacting proteins as well as infrastructure and know-how to process most commercially available multiplex immunoassays.

 

SERVICES

  • Protein profiling of body fluids using customized antibody bead arrays with high sample throughput
    • Study design
    • Antibody selection (from HPA and commercial providers)
    • 384-plex antibody array and direct labeling
    • 100-plex antibody array and direct labeling
    • Data analysis
  • Development of sandwich immunoassays (ELISA type)
  • Protein analysis by multiplexed immunoassays
    • Luminex-based kits (commercial providers)
    • Microchannel-based immunoassays (commercial providers)
  • 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 platform is involved in

  • Screening for potential protein biomarkers with high sample throughput.
  • Building customized antibody bead arrays.
  • Analysis of human body fluids or fluid samples (e.g. lysates)
  • Studies across different disease areas and clinical applications.
  • Assay development for antibody-based protein analysis.

EQUIPMENT

  • Affinity reagents: 50,000  protein array validated antibodies from the Human Protein Atlas project.
  • Instrumentation:  2x FLEXMAP 3D, LX200 and MAGPIX, all from Luminex® and ELLA from ProteinSimple®.
  • Liquid-handling : Tecan FLUENT, 2x Tecan EVO150, CyBio® SELMA.
  • Magnetic bead handler: KingFisher.
  • Plate washer: 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.