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    Enzymatic cleavage of specific dipeptide conjugated with ferrocene as a flexible ultra-sensitive and fast voltammetric assay of matrix metalloproteinase-9 considered a prognostic cancer biomarker in plasma samples

    Journal: Biosens. Bioelectron., Volume 195, JAN 1

    Authors: Nisiewicz, Monika K.; Kowalczyk, Agata; Gajda, Aleksandra; Kasprzak, Artur; Bamburowicz-Klimkowska, Magdalena; Grudzinski, Ireneusz P.; Nowicka, Anna M.

    Organizations: National Science Center Poland [2019/35/B/ST4/02497]; Project TRI-BIO-CHEM; European Social Fund [POWR.03.02.00-00-I007/16-00]

    Keywords: Matrix metalloproteinase-9 activity; Enzymatic peptide cleavage; Peptide-ferrocene conjugate; Voltammetric detection; Sensor; Cancer biomarker detection

    Studies over the last decade have shown that matrix metalloproteinases (MMPs) play a key role in the growth and metastasis of cancer. This zinc-dependent family of endopeptidases is crucial for the degradation of extracellular matrix (ECM), as well as serves as important ECM transducers which have been recognized as early biomarkers for both cancer diagnosis and treatment. In this study, we designed a new type of voltammetric biosensor, composed of a glycine-methionine dipeptide conjugated covalently to ferrocene (Gly-Met-Fc), for fast and ultrasensitive detection of the active form of MMP-9 in plasma samples. The detection was based on specific enzymatic cleavage of the Gly-Met peptide bond, which was monitored by voltammetry and gravimetry measurements. The ferrocene units act as voltammetric visualizers for the detection process. The cysteamine layer directly anchored to the gold surface ensured that the packing density of Gly-Met-Fc in the receptor layer was appropriate for the sensitive detection of MMP-9 in its active form. The developed biosensor was characterized by the widest analytical range (20.10(-6) - 5.0 mu g. mL(-1)) and low detection limit (0.04 pg.mL(-1)). Another valuable feature of the proposed biosensor is that it can be applied directly to the plasma samples without any additional preparation step and thus speeds up the analysis.
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