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Kirichuk, O., Srimasorn, S., Zhang, X. et al. (6 more authors) (2023) Competitive Specific Anchorage of Molecules onto Surfaces: Quantitative Control of Grafting Densities and Contamination by Free Anchors. Langmuir, 39 (50). pp. 18410-18423. ISSN 0743-7463
Abstract
The formation of surfaces decorated with biomacromolecules such as proteins, glycans, or nucleic acids with well-controlled orientations and densities is of critical importance for the design of in vitro models, e.g., synthetic cell membranes and interaction assays. To this effect, ligand molecules are often functionalized with an anchor that specifically binds to a surface with a high density of binding sites, providing control over the presentation of the molecules. Here, we present a method to robustly and quantitatively control the surface density of one or several types of anchor-bearing molecules by tuning the relative concentrations of target molecules and free anchors in the incubation solution. We provide a theoretical background that relates incubation concentrations to the final surface density of the molecules of interest and present effective guidelines toward optimizing incubation conditions for the quantitative control of surface densities. Focusing on the biotin anchor, a commonly used anchor for interaction studies, as a salient example, we experimentally demonstrate surface density control over a wide range of densities and target molecule sizes. Conversely, we show how the method can be adapted to quality control the purity of end-grafted biopolymers such as biotinylated glycosaminoglycans by quantifying the amount of residual free biotin reactant in the sample solution.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2023 The Authors. This is an open access article under the terms of the Creative Commons Attribution License (CC-BY 4.0), which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited. |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) |
Funding Information: | Funder Grant number BBSRC (Biotechnology & Biological Sciences Research Council) BB/T001631/1 Leverhulme Trust RPG-2018-100 |
Depositing User: | Symplectic Publications |
Date Deposited: | 08 Dec 2023 10:59 |
Last Modified: | 22 Mar 2024 10:19 |
Status: | Published |
Publisher: | American Chemical Society |
Identification Number: | 10.1021/acs.langmuir.3c02567 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:206363 |
Available Versions of this Item
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Competitive specific anchorage of molecules onto surfaces: quantitative control of grafting densities and contamination by free anchors. (deposited 05 Jan 2024 14:29)
- Competitive Specific Anchorage of Molecules onto Surfaces: Quantitative Control of Grafting Densities and Contamination by Free Anchors. (deposited 08 Dec 2023 10:59) [Currently Displayed]