RT Journal Article
T1 First bioelectronic immunoplatform for quantitative secretomic analysis of total and metastasis-driven glycosylated haptoglobin
A1 Muñoz San Martín, Cristina
A1 Montero Calle, Ana
A1 Garranzo Asensio, María
A1 Gamella Carballo, Maria
A1 Pérez Ginés, Víctor
A1 Pedrero Muñoz, María
A1 Pingarrón Carrazón, José Manuel
A1 Barderas, Rodrigo
A1 Santos Álvarez, Noemí de los
A1 Lobo Castañón, María Jesús
A1 Campuzano Ruiz, Susana
AB The glycosylation status of proteins is increasingly used as biomarker to improve the reliability in the diagnosis and prognosis of diseases as relevant as cancer. This feeds the need for tools that allow its simple and reliable analysis and are compatible with applicability in the clinic. With this objective in mind, this work reports the frst bioelectronic immunoplatforms described to date for the determination of glycosylated haptoglobin (Hp) and the simultaneous determination of total and glycosylated Hp. The bioelectronic immunoplatform is based on the implementation of non-competitive bioassays using two diferent antibodies or an antibody and a lectin on the surface of commercial magnetic microcarriers. The resulting bioconjugates are labeled with the horseradish peroxidase (HRP) enzyme, and after their magnetic capture on disposable electroplatforms, the amperometric transduction using the H2O2/hydroquinone (HQ) system allows the single or multiple detection. The developed immunoplatform achieves limits of detection (LODs) of 0.07 and 0.46 ng mL−1 for total and glycosylated Hp in bufer solution, respectively. The immunoplatform allows accurate determination using simple and relatively short protocols (approx. 75 min) of total and glycosylated Hp in the secretomes of in vitro–cultured colorectal cancer (CRC) cells with diferent metastatic potentials, which is not feasible, due to lack of sensitivity, by means of some commercial ELISA kits and Western blot methodology.
PB Springer Nature
SN 1618-2642
YR 2022
FD 2022-11-08
LK https://hdl.handle.net/20.500.14352/72692
UL https://hdl.handle.net/20.500.14352/72692
LA eng
NO CRUE-CSIC (Acuerdos Transformativos 2022)
NO Ministerio de Ciencia e Innovación (MICINN)
NO Instituto de Salud Carlos III (ISCIII)/FEDER
NO Comunidad de Madrid
DS Docta Complutense
RD 20 ago 2024