ZAP70 Harmonisation Project

Background

Biomarkers need to fulfil the preliminary steps of technical validation and standardization before proceeding to clinical validation for widespread use. Standardization of technical determination of ZAP-70 expression has proven very challenging. Several factors were evidenced as introducing variability in the results: antibody clone, conjugate, staining strategy, expression of results.

The need for standardization led to the organization of the first international effort on ZAP-70 harmonization in the framework of the ERIC.

Goals

  1. To review the techniques used in the different participating center
  2. To propose a multicentric evaluation of selected procedures
  3. To reach a minimal consensus on good practices
  4. To reach a minimal consensus on the future techncal approach

Participants

Nancy BOECKX Leuven Belgium, Basile STAMATOPOULOS Belgium Sebastien BOETTCHER Kiel Germany, Anne Mette BUHL Copenhagen Denmark, , Paolo GHIA Milano Italy, Rachel IBBOTSON Bournemouth United Kingdom, , Magali LE GARFF-TAVERNIER Paris France, Alison MORILLA London United Kingdom, Ruth PADMORE Ottawa Canada, , Andy RAWSTRON Leeds United Kingdom, Matthias RITGEN Kiel Germany, Medhat. SHEHATA Vienna Austria, Piotr SMOLEWSKI Lodz Poland, Peter STAIB Koln Germany, Michel TICCHIONI Nice France, Neus VILLAMOR Barcelona Spain, Florence CYMBALISTA and Rémi LETESTU Bobigny France.

Approach

The project has gone through 4 previous stages summarized as follows (for more details see completed project)

  1. First stage: a consensus meeting (March 2005)
  2. Second stage: A multicentric evaluation was subsequently designed (July 2006).
  3. Third stage: the electronic-trial (2007)
  4. Fourth stage: design of a new technique (available since January 2010)
    PDF ERIC ZAP-70 staining technique
  5. Fifth stage: evaluation of the new technique (currently ongoing)
    This new technique was further tested among 5 labs on an exchange of fresh samples. A standardization of the settings resulted in similar compensation matrix. Results were homogeneous and consistent with the previous trial. The homogeneity of the results expressed as percentages improved even if the MFI ratio appears more adequate for harmonization, in particular for non-experienced labs.

Time-line

Fifth stage: manuscript preparation mid 2012

Coordination

Pr. Florence Cymbalista, MD, PhD
Bobigny France
E-mail:

Dr Rémi Letestu, MD, PhD
Bobigny France
E-mail: