Servicio de Información Comunitario sobre Investigación y Desarrollo - CORDIS

Stantdardization of MAIPA and lyophilized platelets

Our research & development group of 4 European laboratories was set up in 2001 with the objective to develop simple and affordable assays for HPA platelet typing and antibody detection. One of the first steps of the project consisted in defining a standard protocol for the MAIPA (Monoclonal Antibody Immobilization of Platelet Antigens) assay as a reference protocol.

- The plate-based technique produced very similar results between participants. Good signal/noise ratios were obtained with all HPA specificities investigated. No erroneous results were obtained.
- With the consensus tube method a false-negative reaction was obtained with clinically significant HPA-1 antibodies, anti-GpIbIX antibodies and reactions with HPA-5 system antibodies were less clear. In addition greater variation of results was observed between workshop participants.

This study performed with the same reagents and under the same conditions, showed a lower sensitivity with the tube-based MAIPA when compared with the microplate one. A complementary study comparing three methods (routine tube, consensus tube and microplate method) was performed after the workshop and confirmed the results. Consequently, the group decide to adopt the microplate MAIPA technique as reference technique for the future.

Lyophilized platelets
One of the major drawbacks in the platelet immunology field is the lack of stable reagents to detect allo and autoantibodies. The French team has had the opportunity of getting freezed dried platelets for potential clinical use. We have decided to use them, instead of fresh platelets as is usually done, to check their capacity in the MAIPA test.

Our first step consisted in controlling the surface glycoprotein expression on glycoproteins such as the GPIb, GPIIb, GPIV, CD109 etc.
The level of expression with lyophilized platelets after rehydration turned out to be similar to the one with fresh platelets. The same in the MAIPA tests, lyophilized platelets gave results similar to the results obtained with fresh platelets. This new reagent is being validated by other laboratories (CR2, British team /CR4 German team and CR5 Swiss team) to check that all types of allo and autoantibodies can be detected.

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