2 research outputs found
Disposable amperometric immunosensor for the determination of the Eâcadherin tumor suppressor protein in cancer cells and human tissues
This paper describes the results obtained in the development of the first electrochemical immunosensor described to date for the detection of Eâcadherin (Eâcad) protein, a relevant biomarker of prognosis and metastasis in cancer, based on the use of magnetic microcarriers (MBs) and amperometric transduction at screenâprinted carbon electrodes (SPCEs). Thus, the determination of Eâcad protein involved the use of two specific antibodies against this protein (one of them labelled with HRP) in a sandwich configuration onto HOOCâMBs. The magnetic bioconjugates were captured onto SPCEs and the amperometric transduction was performed using the H2O2/hydroquinone (HQ) system. Under optimal conditions, this bioplatform demonstrated a wide linear concentration range (0.50â25â
ngâmLâ1) and a detection limit as low as 0.16â
ngâmLâ1, well below the optimal cutâoff level for the Eâcad protein (defined as 10,000â
ngâmLâ1 for soluble Eâcad levels in serum). The developed sensor also showed a good reproducibility among measurements with seven different sensors constructed in the same manner (RSD, 5.4â%), stability for more than 15â
days and good specificity towards other proteins commonly found on biological samples. The applicability of this simple handling bioplatform for the direct determination of this protein in cell lysates with different metastatic potential and extracts from paraffinedâembedded human colorectal cancer tissues of different grade were also demonstrated.The financial support of the Spanish Ministerio de EconomĂa y Competitividad (CTQ2015â64402âC2â1âR Project) and the NANOAVANSENS Program from the Comunidad de Madrid (S2013/MTâ3029 Project) are gratefully acknowledged. C.M.S.M. was supported by a contract of the S2013/MTâ3029 Project. R.B. acknowledges the financial support of the PI17CIII/00045 grant from the AESâISCIII program. M.G.A. was supported by a contract of the Programa Operativo de Empleo Juvenil y la Iniciativa de Empleo Juvenil (YEI) with the participation of the ConsejerĂa de EducaciĂłn, Juventud y Deporte de la Comunidad de Madrid y del Fondo Social Europeo.Peer reviewe