WNT-mediated signaling relay in stem cells and oncogenesis - from basic biology to applications

QCM-based WNT binding assay to quantitatively profile WNT-receptor interactions

Experienced researcher 1 (ER1) project

Supervision: Teodor Aastrup

Lab webpage:  http://www.attana.com

Aim:
Technical development of quantitative WNT-binding assays and adaption to semi-high throughput format.

Methodology:
ER1 will use the quartz crystal microbalance (QCM) technique, which is highly powerful to monitor adsorption/binding events to surfaces, and is increasingly applied in biorecognition analysis. The technique, based on piezoelectric properties, offers label-free binding measurements in real-time, with high sensitivity and reliability. ER1 will employ variable chip surfaces, which are suitable for binding proteins, membranes and even living/fixed cells.

Collaborators:
Schulte, Sunahara, Maurice, Pepscan

Project goals:
ER1 will develop an HTS-compatible WNT binding assay for affinity determination and kinetic analysis of agonist-receptor interactions, suitable for drug discovery.

Risk assessment and contingency plans:
Methodology and technical platforms were developed by Attana and are routinely used for biorecognition studies, such as ligand-receptor interactions. The challenge is the lipophilic character of WNTs, which will be addressed by evaluation of whole cells, cell membrane preparations, and FZD-HDL particles (collaboration Sunahara, international secondment: Schulte), offering the system different amounts of unspecific binding sites. For proof-of-concept studies, FZD antibodies as well as FZD ligands without lipid modification (collaboration Maurice, private sector secondment: Pepscan) will be employed.

Key publications:

  1. Pei Z, Saint-Guirons J, Käck C, Ingemarsson B, Aastrup T. (2012) Real-time analysis of the carbohydrates on cell surfaces using a QCM biosensor: a lectin-based approach. Biosens Bioelectron. 35(1):200-5.
  2. Mahon E, Aastrup T, Barboiu M. (2010) Multivalent recognition of lectins by glyconanoparticle systems. Chem Commun. 46(30):5491-3.
  3. Mahon E, Aastrup T, Barboiu M. (2010) Dynamic glycovesicle systems for amplified QCM detection of carbohydrate-lectin multivalent biorecognition. Chem Commun. 46(14):2441-3