Reactive Thin Polymer Films as Platforms for the Immobilization of Biomolecules

Chuan Liang Feng, Zheng Zhang, Renate Förch, Wolfgang Knoll, G. Julius Vancso, Holger Schönherr

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Spin-coated thin films of poly(N-hydroxysuccinimidyl methacrylate) (PNHSMA) on oxidized silicon and gold surfaces were investigated as reactive layers for obtaining platforms for biomolecule immobilization with high molecular loading. The surface reactivity of PNHSMA films in coupling reactions with various primary amines, including amine-terminated poly(ethylene glycol) (PEG-NH2) and fluoresceinamine, was determined by Fourier transform infrared (FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), fluorescence microscopy, and ellipsometry measurements, respectively. The rate constants of PEG-NH2 attachment on the PNHSMA films were found to be significantly increased compared to the coupling on self-assembled monolayers (SAMs) of 11,11‘-dithiobis(N-hydroxysuccinimidylundecanoate) (NHS-C10) on gold under the same conditions. More significantly, the PEG loading observed was about 3 times higher for the polymer thin films. These data indicate that the coupling reactions are not limited to the very surface of the polymer films, but proceed into the near-surface regions of the films. PNHSMA films were shown to be stable in contact with aqueous buffer; the swelling analysis, as performed by atomic force microscopy (AFM), indicated a film thickness independent swelling of 2 nm. An increased loading was also observed by surface plasmon resonance for the covalent immobilization of amino-functionalized probe DNA. Hybridization of fluorescently labeled target DNA was successfully detected by fluorescence microscopy and surface plasmon resonance enhanced fluorescence spectroscopy (SPFS), thereby demonstrating that thin films of PNHSMA comprise an attractive and simple platform for the immobilization of biomolecules with high densities.
Original languageEnglish
Pages (from-to)3243-3251
Number of pages9
JournalBiomacromolecules
Volume6
Issue number6
DOIs
Publication statusPublished - 2005

Fingerprint

Biomolecules
Polymer films
Polyethylene glycols
Thin films
Fluorescence microscopy
Surface plasmon resonance
Swelling
Amines
DNA
Gold
Ellipsometry
Fluorescence spectroscopy
Self assembled monolayers
Fourier transform infrared spectroscopy
DNA Probes
Film thickness
Silicon
Rate constants
Atomic force microscopy
X ray photoelectron spectroscopy

Cite this

Feng, Chuan Liang ; Zhang, Zheng ; Förch, Renate ; Knoll, Wolfgang ; Vancso, G. Julius ; Schönherr, Holger. / Reactive Thin Polymer Films as Platforms for the Immobilization of Biomolecules. In: Biomacromolecules. 2005 ; Vol. 6, No. 6. pp. 3243-3251.
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abstract = "Spin-coated thin films of poly(N-hydroxysuccinimidyl methacrylate) (PNHSMA) on oxidized silicon and gold surfaces were investigated as reactive layers for obtaining platforms for biomolecule immobilization with high molecular loading. The surface reactivity of PNHSMA films in coupling reactions with various primary amines, including amine-terminated poly(ethylene glycol) (PEG-NH2) and fluoresceinamine, was determined by Fourier transform infrared (FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), fluorescence microscopy, and ellipsometry measurements, respectively. The rate constants of PEG-NH2 attachment on the PNHSMA films were found to be significantly increased compared to the coupling on self-assembled monolayers (SAMs) of 11,11‘-dithiobis(N-hydroxysuccinimidylundecanoate) (NHS-C10) on gold under the same conditions. More significantly, the PEG loading observed was about 3 times higher for the polymer thin films. These data indicate that the coupling reactions are not limited to the very surface of the polymer films, but proceed into the near-surface regions of the films. PNHSMA films were shown to be stable in contact with aqueous buffer; the swelling analysis, as performed by atomic force microscopy (AFM), indicated a film thickness independent swelling of 2 nm. An increased loading was also observed by surface plasmon resonance for the covalent immobilization of amino-functionalized probe DNA. Hybridization of fluorescently labeled target DNA was successfully detected by fluorescence microscopy and surface plasmon resonance enhanced fluorescence spectroscopy (SPFS), thereby demonstrating that thin films of PNHSMA comprise an attractive and simple platform for the immobilization of biomolecules with high densities.",
author = "Feng, {Chuan Liang} and Zheng Zhang and Renate F{\"o}rch and Wolfgang Knoll and Vancso, {G. Julius} and Holger Sch{\"o}nherr",
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Feng, CL, Zhang, Z, Förch, R, Knoll, W, Vancso, GJ & Schönherr, H 2005, 'Reactive Thin Polymer Films as Platforms for the Immobilization of Biomolecules' Biomacromolecules, vol. 6, no. 6, pp. 3243-3251. https://doi.org/10.1021/bm050247u

Reactive Thin Polymer Films as Platforms for the Immobilization of Biomolecules. / Feng, Chuan Liang; Zhang, Zheng; Förch, Renate; Knoll, Wolfgang; Vancso, G. Julius; Schönherr, Holger.

In: Biomacromolecules, Vol. 6, No. 6, 2005, p. 3243-3251.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - Reactive Thin Polymer Films as Platforms for the Immobilization of Biomolecules

AU - Feng, Chuan Liang

AU - Zhang, Zheng

AU - Förch, Renate

AU - Knoll, Wolfgang

AU - Vancso, G. Julius

AU - Schönherr, Holger

PY - 2005

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N2 - Spin-coated thin films of poly(N-hydroxysuccinimidyl methacrylate) (PNHSMA) on oxidized silicon and gold surfaces were investigated as reactive layers for obtaining platforms for biomolecule immobilization with high molecular loading. The surface reactivity of PNHSMA films in coupling reactions with various primary amines, including amine-terminated poly(ethylene glycol) (PEG-NH2) and fluoresceinamine, was determined by Fourier transform infrared (FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), fluorescence microscopy, and ellipsometry measurements, respectively. The rate constants of PEG-NH2 attachment on the PNHSMA films were found to be significantly increased compared to the coupling on self-assembled monolayers (SAMs) of 11,11‘-dithiobis(N-hydroxysuccinimidylundecanoate) (NHS-C10) on gold under the same conditions. More significantly, the PEG loading observed was about 3 times higher for the polymer thin films. These data indicate that the coupling reactions are not limited to the very surface of the polymer films, but proceed into the near-surface regions of the films. PNHSMA films were shown to be stable in contact with aqueous buffer; the swelling analysis, as performed by atomic force microscopy (AFM), indicated a film thickness independent swelling of 2 nm. An increased loading was also observed by surface plasmon resonance for the covalent immobilization of amino-functionalized probe DNA. Hybridization of fluorescently labeled target DNA was successfully detected by fluorescence microscopy and surface plasmon resonance enhanced fluorescence spectroscopy (SPFS), thereby demonstrating that thin films of PNHSMA comprise an attractive and simple platform for the immobilization of biomolecules with high densities.

AB - Spin-coated thin films of poly(N-hydroxysuccinimidyl methacrylate) (PNHSMA) on oxidized silicon and gold surfaces were investigated as reactive layers for obtaining platforms for biomolecule immobilization with high molecular loading. The surface reactivity of PNHSMA films in coupling reactions with various primary amines, including amine-terminated poly(ethylene glycol) (PEG-NH2) and fluoresceinamine, was determined by Fourier transform infrared (FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), fluorescence microscopy, and ellipsometry measurements, respectively. The rate constants of PEG-NH2 attachment on the PNHSMA films were found to be significantly increased compared to the coupling on self-assembled monolayers (SAMs) of 11,11‘-dithiobis(N-hydroxysuccinimidylundecanoate) (NHS-C10) on gold under the same conditions. More significantly, the PEG loading observed was about 3 times higher for the polymer thin films. These data indicate that the coupling reactions are not limited to the very surface of the polymer films, but proceed into the near-surface regions of the films. PNHSMA films were shown to be stable in contact with aqueous buffer; the swelling analysis, as performed by atomic force microscopy (AFM), indicated a film thickness independent swelling of 2 nm. An increased loading was also observed by surface plasmon resonance for the covalent immobilization of amino-functionalized probe DNA. Hybridization of fluorescently labeled target DNA was successfully detected by fluorescence microscopy and surface plasmon resonance enhanced fluorescence spectroscopy (SPFS), thereby demonstrating that thin films of PNHSMA comprise an attractive and simple platform for the immobilization of biomolecules with high densities.

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DO - 10.1021/bm050247u

M3 - Article

VL - 6

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EP - 3251

JO - Biomacromolecules

JF - Biomacromolecules

SN - 1525-7797

IS - 6

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