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ELISA validation approach for the detection of anti-saccharomyces cerevisiae antibodies in patients treated with biopharmaceutical heberprot-P®


Journal of Analytical & Pharmaceutical Research
Maylin Pérez-Bernal,1 Carlos Hernández,Magali Delgado,1 Mabel Izquierdo,2 Ileana Rosales,2 Enrique Pérez1

Abstract

This  work  describes  the  validation  of  an  enzyme-linked  immunosorbent  assay  (ELISA)  for  detection  of  anti-Saccharomyces  cerevisiae  antibodies  (ASCA)  in  diabetic  patients  with  foot  ulcers,  after  the  treatment  with  Heberprot-P®.  Validation  followed  regulatory  guidelines  of  US  FDA  and  European  Medicine  Agency.  Minimum  required  dilution  of samples and quality controls were defined using pools of sera from diabetic patients and from healthy donors. Parameters such as cut point, specificity, precision, selectivity, robustness and sample stability were analyzed. The repeatability and intermediate precision percent ranged between 7.93-10.61% and 7.93-11.43 %, respectively, indicating low intra- and inter-assay variation. The specificity was proved by background noise suppression, reaching 100% of inhibition as strong criterion for the specificity of the immunoassay. The  validated  ELISA  is  a  reliable  tool  for  ASCA  detection  in  human  serum  after  the  administration of Heberprot-P®, in order to find immunological reactions associated with latent contamination by host cell proteins from Saccharomyces cerevisiae.

Keywords

anti-Saccharomyces cerevisiae antibodies, diabetic patients, ELISA validation, serum samples

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