First bioelectronic immunoplatform for quantitative secretomic analysis of total and metastasis-driven glycosylated haptoglobin



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Muñoz San Martín, Cristina and Montero Calle, Ana and Garranzo Asensio, María and Gamella Carballo, Maria and Pérez Ginés, Víctor and Pedrero Muñoz, María and Pingarrón Carrazón, José Manuel and Barderas, Rodrigo and Santos Álvarez, Noemí de los and Lobo Castañón, María Jesús and Campuzano Ruiz, Susana (2022) First bioelectronic immunoplatform for quantitative secretomic analysis of total and metastasis-driven glycosylated haptoglobin. Analytical and Bioanalytical Chemistry . ISSN 1618-2642

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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.

Item Type:Article
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CRUE-CSIC (Acuerdos Transformativos 2022)

Uncontrolled Keywords:Glycosylated haptoglobin; Amperometry; Multiplexed immunoplatform; Secretome; Metastatic CRC cells
Subjects:Sciences > Chemistry > Analytic chemistry
ID Code:75678
Deposited On:24 Nov 2022 11:59
Last Modified:24 Nov 2022 12:02

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