Antifouling PEG Coatings by Thiol‐Acrylate Conjugate Addition Reactions for Generation of Protein Patterns via Photobleaching‐Induced Protein Binding (PiPB) BioBitmaps Using Maskless Projection Lithography
Abstract Photobleaching‐induced protein binding (PiPB) is a light‐based molecular patterning technique that is introduced as Protein Bitmaps. This technique has significant applications in immunoassays and cell‐substrate interactions. However, commonly used active surfaces, bovine serum albumin (BSA...
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Wiley-VCH
2025-06-01
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| Series: | Advanced Materials Interfaces |
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| Online Access: | https://doi.org/10.1002/admi.202500198 |
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| author | Ali Usama Evelin Schäfer Pang Zhu Qingchuan Song Alexander Weißbach Dorothea Helmer Joachim Wollschläger Changjiang You Martin Steinhart Pegah Pezeshkpour Peilong Hou Bastian E. Rapp |
| author_facet | Ali Usama Evelin Schäfer Pang Zhu Qingchuan Song Alexander Weißbach Dorothea Helmer Joachim Wollschläger Changjiang You Martin Steinhart Pegah Pezeshkpour Peilong Hou Bastian E. Rapp |
| author_sort | Ali Usama |
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| description | Abstract Photobleaching‐induced protein binding (PiPB) is a light‐based molecular patterning technique that is introduced as Protein Bitmaps. This technique has significant applications in immunoassays and cell‐substrate interactions. However, commonly used active surfaces, bovine serum albumin (BSA) coatings prepared via physical adsorption, are prone to desorption or displacement by biomolecules with higher substrate affinities, limiting their stability under complex conditions over several days. To address this, covalently bound high‐density antifouling polyethylene glycol (PEG) monolayers are developed by solvent‐free coupling of cost‐effective homo‐bifunctional PEGs (acrylate‐PEG‐acrylate, PEGDA) to glass/glass‐type surfaces silanized with (3‐mercaptopropyl) trimethoxysilane (MPTMS) using thiol‐acrylate conjugate addition reactions at room temperature, resulting in stable antifouling PEGDA surfaces with terminal‐acrylates as free radical acceptors for PiPB. Non‐specific protein binding on PEGDA‐modified surfaces is evaluated using reflectometric interference spectroscopy (RIfS), showing superior antifouling performance compared to BSA‐coated surfaces against avidin and comparable performance against streptavidin and BSA. Furthermore, PiPB with fluorescein‐5‐biotin conjugate (F5B) is carried out on PEGDA‐modified surfaces, performed using a custom‐built digital mirror device (DMD)‐based lithography system, confirming the suitability of PEGDA‐modified surfaces for biomolecule immobilization. The method presented for PEGDA coating preparation has the potential to broaden the applicability of PiPB, particularly using DMD‐based devices, in biomedical and surface engineering fields. |
| format | Article |
| id | doaj-art-cf0e4f7a82b944e4a2b2db28a68b7088 |
| institution | Matheson Library |
| issn | 2196-7350 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Wiley-VCH |
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| series | Advanced Materials Interfaces |
| spelling | doaj-art-cf0e4f7a82b944e4a2b2db28a68b70882025-06-30T08:53:44ZengWiley-VCHAdvanced Materials Interfaces2196-73502025-06-011212n/an/a10.1002/admi.202500198Antifouling PEG Coatings by Thiol‐Acrylate Conjugate Addition Reactions for Generation of Protein Patterns via Photobleaching‐Induced Protein Binding (PiPB) BioBitmaps Using Maskless Projection LithographyAli Usama0Evelin Schäfer1Pang Zhu2Qingchuan Song3Alexander Weißbach4Dorothea Helmer5Joachim Wollschläger6Changjiang You7Martin Steinhart8Pegah Pezeshkpour9Peilong Hou10Bastian E. Rapp11Laboratory of Process Technology NeptunLab Department of Microsystems Engineering (IMTEK) University of Freiburg Georges‐Köhler‐Allee 103 79110 Freiburg im Breisgau GermanyDepartment of Biology/Chemistry and Center for Cellular Nanoanalytics (CellNanOs) Universität Osnabrück Barbarastr. 7 49076 Osnabrück GermanyLaboratory of Process Technology NeptunLab Department of Microsystems Engineering (IMTEK) University of Freiburg Georges‐Köhler‐Allee 103 79110 Freiburg im Breisgau GermanyLaboratory of Process Technology NeptunLab Department of Microsystems Engineering (IMTEK) University of Freiburg Georges‐Köhler‐Allee 103 79110 Freiburg im Breisgau GermanyDepartment of Mathematics/Informatics/Physics Universität Osnabrück Barbarastr. 7 49076 Osnabrück GermanyLaboratory of Process Technology NeptunLab Department of Microsystems Engineering (IMTEK) University of Freiburg Georges‐Köhler‐Allee 103 79110 Freiburg im Breisgau GermanyDepartment of Mathematics/Informatics/Physics Universität Osnabrück Barbarastr. 7 49076 Osnabrück GermanyDepartment of Biology/Chemistry and Center for Cellular Nanoanalytics (CellNanOs) Universität Osnabrück Barbarastr. 7 49076 Osnabrück GermanyDepartment of Biology/Chemistry and Center for Cellular Nanoanalytics (CellNanOs) Universität Osnabrück Barbarastr. 7 49076 Osnabrück GermanyLaboratory of Process Technology NeptunLab Department of Microsystems Engineering (IMTEK) University of Freiburg Georges‐Köhler‐Allee 103 79110 Freiburg im Breisgau GermanyLaboratory of Process Technology NeptunLab Department of Microsystems Engineering (IMTEK) University of Freiburg Georges‐Köhler‐Allee 103 79110 Freiburg im Breisgau GermanyLaboratory of Process Technology NeptunLab Department of Microsystems Engineering (IMTEK) University of Freiburg Georges‐Köhler‐Allee 103 79110 Freiburg im Breisgau GermanyAbstract Photobleaching‐induced protein binding (PiPB) is a light‐based molecular patterning technique that is introduced as Protein Bitmaps. This technique has significant applications in immunoassays and cell‐substrate interactions. However, commonly used active surfaces, bovine serum albumin (BSA) coatings prepared via physical adsorption, are prone to desorption or displacement by biomolecules with higher substrate affinities, limiting their stability under complex conditions over several days. To address this, covalently bound high‐density antifouling polyethylene glycol (PEG) monolayers are developed by solvent‐free coupling of cost‐effective homo‐bifunctional PEGs (acrylate‐PEG‐acrylate, PEGDA) to glass/glass‐type surfaces silanized with (3‐mercaptopropyl) trimethoxysilane (MPTMS) using thiol‐acrylate conjugate addition reactions at room temperature, resulting in stable antifouling PEGDA surfaces with terminal‐acrylates as free radical acceptors for PiPB. Non‐specific protein binding on PEGDA‐modified surfaces is evaluated using reflectometric interference spectroscopy (RIfS), showing superior antifouling performance compared to BSA‐coated surfaces against avidin and comparable performance against streptavidin and BSA. Furthermore, PiPB with fluorescein‐5‐biotin conjugate (F5B) is carried out on PEGDA‐modified surfaces, performed using a custom‐built digital mirror device (DMD)‐based lithography system, confirming the suitability of PEGDA‐modified surfaces for biomolecule immobilization. The method presented for PEGDA coating preparation has the potential to broaden the applicability of PiPB, particularly using DMD‐based devices, in biomedical and surface engineering fields.https://doi.org/10.1002/admi.202500198antifouling PEG coatingsDMD‐based maskless projection lithographyphotobleachingprotein bitmapsthiol‐acrylate conjugate addition reactions |
| spellingShingle | Ali Usama Evelin Schäfer Pang Zhu Qingchuan Song Alexander Weißbach Dorothea Helmer Joachim Wollschläger Changjiang You Martin Steinhart Pegah Pezeshkpour Peilong Hou Bastian E. Rapp Antifouling PEG Coatings by Thiol‐Acrylate Conjugate Addition Reactions for Generation of Protein Patterns via Photobleaching‐Induced Protein Binding (PiPB) BioBitmaps Using Maskless Projection Lithography Advanced Materials Interfaces antifouling PEG coatings DMD‐based maskless projection lithography photobleaching protein bitmaps thiol‐acrylate conjugate addition reactions |
| title | Antifouling PEG Coatings by Thiol‐Acrylate Conjugate Addition Reactions for Generation of Protein Patterns via Photobleaching‐Induced Protein Binding (PiPB) BioBitmaps Using Maskless Projection Lithography |
| title_full | Antifouling PEG Coatings by Thiol‐Acrylate Conjugate Addition Reactions for Generation of Protein Patterns via Photobleaching‐Induced Protein Binding (PiPB) BioBitmaps Using Maskless Projection Lithography |
| title_fullStr | Antifouling PEG Coatings by Thiol‐Acrylate Conjugate Addition Reactions for Generation of Protein Patterns via Photobleaching‐Induced Protein Binding (PiPB) BioBitmaps Using Maskless Projection Lithography |
| title_full_unstemmed | Antifouling PEG Coatings by Thiol‐Acrylate Conjugate Addition Reactions for Generation of Protein Patterns via Photobleaching‐Induced Protein Binding (PiPB) BioBitmaps Using Maskless Projection Lithography |
| title_short | Antifouling PEG Coatings by Thiol‐Acrylate Conjugate Addition Reactions for Generation of Protein Patterns via Photobleaching‐Induced Protein Binding (PiPB) BioBitmaps Using Maskless Projection Lithography |
| title_sort | antifouling peg coatings by thiol acrylate conjugate addition reactions for generation of protein patterns via photobleaching induced protein binding pipb biobitmaps using maskless projection lithography |
| topic | antifouling PEG coatings DMD‐based maskless projection lithography photobleaching protein bitmaps thiol‐acrylate conjugate addition reactions |
| url | https://doi.org/10.1002/admi.202500198 |
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