2026-04-29T04:10:24Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/159052025-06-13T10:05:26Zcom_20.500.12105_15322com_20.500.12105_2051com_20.500.12105_2052col_20.500.12105_16990col_20.500.12105_19617
Repisalud
author
Muñoz-San Martín, Cristina
author
Montero-Calle, Ana Maria
author
Garranzo-Asensio, Maria
author
Gamella, Maria
author
Pérez-Ginés, Víctor
author
Pedrero, María
author
Pingarrón, José M
author
Barderas Manchado, Rodrigo
author
de-Los-Santos-Álvarez, Noemí
author
Lobo-Castañón, María Jesús
author
Campuzano, Susana
funder
Consejo Superior de Investigaciones Científicas (España)
funder
Conferencia de Rectores de las Universidades Españolas
funder
Ministerio de Ciencia e Innovación (España)
funder
Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)
funder
Comunidad de Madrid (España)
funder
Instituto de Salud Carlos III
2023-04-27T07:56:25Z
2023-04-27T07:56:25Z
2023-05
Anal Bioanal Chem. 2023 May;415(11):2045-2057.
http://hdl.handle.net/20.500.12105/15905
36344668
10.1007/s00216-022-04397-6
1618-2650
Analytical and bioanalytical chemistry
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 first 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 different 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 buffer 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 different metastatic potentials, which is not feasible, due to lack of sensitivity, by means of some commercial ELISA kits and Western blot methodology.
eng
Amperometry
Glycosylated haptoglobin
Metastatic CRC cells
Multiplexed immunoplatform
Secretome
First bioelectronic immunoplatform for quantitative secretomic analysis of total and metastasis-driven glycosylated haptoglobin
research article
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FirstBioelectronicImmunoplatformQuantitative_2023.pdf
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Supplementary_FirstBioelectronicImmunoplatformQuantitative_2023.pdf
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FirstBioelectronicImmunoplatformQuantitative_2023.pdf.txt
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Supplementary_FirstBioelectronicImmunoplatformQuantitative_2023.pdf.txt