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Density Control over MBD2 Receptor-Coated Surfaces Provides Superselective Binding of Hypermethylated DNA
Using the biomarker hypermethylated DNA (hmDNA) for cancer detection requires a pretreatment to isolate or concentrate hmDNA from nonmethylated DNA. Affinity chromatography using a methyl binding domain-2 (MBD2) protein can be used, but the relatively low enrichment selectivity of MBD2 limits its cl...
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Published in: | ACS applied materials & interfaces 2022-09, Vol.14 (36), p.40579-40589 |
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creator | Kolkman, Ruben W. Michel-Souzy, Sandra Wasserberg, Dorothee Segerink, Loes I. Huskens, Jurriaan |
description | Using the biomarker hypermethylated DNA (hmDNA) for cancer detection requires a pretreatment to isolate or concentrate hmDNA from nonmethylated DNA. Affinity chromatography using a methyl binding domain-2 (MBD2) protein can be used, but the relatively low enrichment selectivity of MBD2 limits its clinical applicability. Here, we developed a superselective, multivalent, MBD2-coated platform to improve the selectivity of hmDNA enrichment. The multivalent platform employs control over the MBD2 surface receptor density, which is shown to strongly affect the binding of DNA with varying degrees of methylation, improving both the selectivity and the affinity of DNAs with higher numbers of methylation sites. Histidine-10-tagged MBD2 was immobilized on gold surfaces with receptor density control by tuning the amount of nickel nitrilotriacetic acid (NiNTA)-functionalized thiols in a thiol-based self-assembled monolayer. The required MBD2 surface receptor densities for DNA surface binding decreases for DNA with higher degrees of methylation. Both higher degrees of superselectivity and surface coverages were observed upon DNA binding at increasing methylation levels. Adopting the findings of this study into hmDNA enrichment of clinical samples has the potential to become more selective and sensitive than current MBD2-based methods and, therefore, to improve cancer diagnostics. |
doi_str_mv | 10.1021/acsami.2c09641 |
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Both higher degrees of superselectivity and surface coverages were observed upon DNA binding at increasing methylation levels. Adopting the findings of this study into hmDNA enrichment of clinical samples has the potential to become more selective and sensitive than current MBD2-based methods and, therefore, to improve cancer diagnostics.</description><identifier>ISSN: 1944-8244</identifier><identifier>EISSN: 1944-8252</identifier><identifier>DOI: 10.1021/acsami.2c09641</identifier><identifier>PMID: 36052432</identifier><language>eng</language><publisher>American Chemical Society</publisher><subject>Biological and Medical Applications of Materials and Interfaces</subject><ispartof>ACS applied materials & interfaces, 2022-09, Vol.14 (36), p.40579-40589</ispartof><rights>2022 The Authors. Published by American Chemical Society</rights><rights>2022 The Authors. 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The required MBD2 surface receptor densities for DNA surface binding decreases for DNA with higher degrees of methylation. Both higher degrees of superselectivity and surface coverages were observed upon DNA binding at increasing methylation levels. 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subjects | Biological and Medical Applications of Materials and Interfaces |
title | Density Control over MBD2 Receptor-Coated Surfaces Provides Superselective Binding of Hypermethylated DNA |
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