A MALDI-MS sensing chip prepared by non-covalent assembly for quantitation of acid phosphatase
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MALDI-MS sensing chip prepared by non-covalent assembly for quantitation of acid phosphatase 1†
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Qiulin Ma , Yunlong Chen , Nan Feng , Feng Yan & Huangxian Ju 1
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State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University,
Nanjing 210023, China; Department Clinical Laboratory, Affiliated Cancer Hospital of Nanjing Medical University, Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, Nanjing 210009, China Received June 20, 2020; accepted August 14, 2020; published online September 28, 2020
A novel sensing chip was designed for MALDI-MS quantitation of acid phosphatase (ACP). The ACP sensing chip was constructed through non-covalent interaction of streptavidin and biotin for the assembly of biotinylated peptide substrate on biotinylated polyethylene-glycol (PEG) modified indium-tin oxide (ITO) slide. In the presence of ACP, the peptide substrate was dephosphorylated under acidic condition to generate a new mass signal. The quantitative assay of ACP was achieved with the mass signal ratio of product to the sum of product and left peptide substrate. Under optimal detection conditions, the ratio was linearly correlated with the concentration of ACP in the range of 0.05–12 g/L with a detection limit (LOD) of 0.04 g/L. The designed ACP sensing chip has been used to analyze ACP in complex clinical samples, which exhibited high selectivity, good repeatability, and admirably anti-interference ability. This work further demonstrates the concept of MS sensing and the application of MALDI-MS in quantitative analysis, and provides a convenient method for the quantitation of proteases in clinical diagnosis. acid phosphatase, sensing chip, MALDI-MS quantitation, proteases, mass signal Citation:
Ma Q, Chen Y, Feng N, Yan F, Ju H. A MALDI-MS sensing chip prepared by non-covalent assembly for quantitation of acid phosphatase. Sci China Chem, 2020, 63, https://doi.org/10.1007/s11426-020-9850-3
1 Introduction Acid phosphatase (ACP) is a ubiquitous natural protease found in most of animals and plants, especially in mammalian tissues and fluids, such as prostate and liver [1,2]. It can remove a phosphate group from its substrate into a phosphate ion so as to catalyze the hydrolysis of orthophosphate monoesters and transphosphorylation reactions under acidic condition [3], which plays an essential role in many physiological processes. According to clinical studies, 30%– 50% of ACP in normal male serum comes from prostate, while ACP in female serum comes from liver, red blood cells †These authors contributed equally to this work. *Corresponding author (email: [email protected])
and platelets. ACP concentration in healthy human serum is about 0–9 U/L. The abnormal expression of ACP in human serum may indicate the occurrence of some diseases. When prostate cancer occurs or metastasizes, the content of ACP in blood is significantly increased, which demonstrates its great value for the diagnosis, curative effect and prognosis monitoring of advanc
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