Développement de la détection de l’hematide

Code: R09B2JD

In addition to the increasing number of different recombinant erythropoietins (rhEPO) that are present on the market at the present time, peptide-mimetics of these hormones producing identical effects but presenting unrelated molecular structures will be soon commercialized. Due to this difference in structure, such mimetics cannot be detected by the test used for anti-doping control of rhEPO. Specific methods have to be developed.

One of these peptide mimetics, peginesatide (formerly known as Hematide™), produced by the biopharmaceutical company Affymax is now investigated in Phase 3 trials for the potential treatment of anemia associated with chronic kidney disease (CKD). This drug figures on the WADA list of prohibited substances in sport.

Affymax Company is collaborating with WADA in order that anti-doping laboratories have a test to detect Hematide at their disposal.

Affymax has developed different antibodies directed against Hematide and has tested these antibodies in two different methods, ELISA and immunoblot following SDS electrophoresis.

This project was a collaborative study including Affymax and the anti-doping laboratories of Lausanne (Switzerland) and Châtenay-Malabry (France) and was intended to validate the two different methods proposed by Affymax in two different laboratories.

The aim of the project was to investigate, eventually improve and provide a full validation of the two methods in order that they can be used for detection of the drug in blood samples as screening(ELISA) and confirmation (SDS electrophoresis) tests.

Investigations of ELISA and SDS electrophoresis for eventual improvements were especially performed by the Swiss and French laboratories respectively. Validation of both methods was performed by both laboratories.

Main findings

After some modifications of the SDS electrophoresis method, our laboratory has fully validated both this method and the ELISA method as transmitted by the laboratory of Lausanne.

Our investigations have shown that both methods are specific and robust and can thus be applied for anti-doping control of Hematide using either plasma or serum samples.

Their limit of detection are similar (0.1 ng/mL for ELISA and <0.25 ng/mL for SDS electrophoresis). They have been used in a blind test performed on blood samples from a clinical trial and provided by Affymax to our laboratory. Neither false negative nor false positive results have been observed.

The window of detection was 21 to 28 days following an injection of 0.05 mg/kg (IV).