Protein immobilization and detection on laser processed polystyrene surfaces

Journal Article · Thu Sep 15 00:00:00 EDT 2011 · Journal of Applied Physics

DOI:https://doi.org/10.1063/1.3627160· OSTI ID:22036727

Sarantopoulou, Evangelia; Kollia, Zoe; Palles, Dimitrios; Spyropoulos-Antonakakis, Nikolaos; Cefalas, Alkiviadis-Constantinos ^[1]; Petrou, Panagiota S; Kakabakos, Sotirios ^[2]

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1. National Hellenic Research Foundation, Theoretical and Physical Chemistry Institute, 48 Vassileos Constantinou Avenue, Athens 11635 (Greece)
2. N.C.S.R. ''Demokritos'', Institute of Radioisotopes and Radiodiagnostic Products, Immunoassay/Immunosensors Lab., Patriarchou Gregoriou Str, Aghia Paraskevi, Athens 15310 (Greece)

The bovine serum albumin (BSA)-polystyrene (PS) interface layer is laser photo activated at 157 nm for site selective multiple target-protein immobilization. The 5-15 nm photon induced interface layer has different chemical, wetting, and stiffness properties than the PS photon processed surface. The irradiated areas exhibit target-protein binding, followed by localized probe-target protein detection. The photon induced chemical modification of the BSA-PS interface layer is identified by: (1) Morphological, imaging, and analysis of surface parameters with atomic force microscopy, (2) spectroscopic shift (4 cm{sup -1}), of the amide I group and formation of new C=N, NH{sub 2}, C-O, C=O, and O-C=O groups following irradiation, identified with attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, and (3) the different hydrophilic/hydrophobic and force-distance response of the bare PS and BSA-PS surfaces. Near field edge diffraction (Fresnel) fluorescence imaging specifies the threshold photon energy and the fluence required to optically detect the protein binding on the photon induced BSA-PS interface layer. By approximating the Fresnel integrals with analytical functions, the threshold photon energy and the fluence are expressed as the sum of zero, first, and second order harmonic terms of two characteristic diffracted modes and they are specified to be 8.73x10{sup -9} Jand623 J m{sup -2}, respectively. Furthermore, a bioarray of three probe-target proteins is fabricated with 1.5 {mu}m spatial resolution using a 157 nm laser microstepper. The methodology eliminates the use of intermediate polymer layers between the blocking BSA protein and the PS substrate in bioarray fabrication.

• OSTI ID: 22036727
• Journal Information: Journal of Applied Physics, Vol. 110, Issue 6; Other Information: (c) 2011 American Institute of Physics; Country of input: International Atomic Energy Agency (IAEA); ISSN 0021-8979
• Country of Publication: United States
• Language: English

Related Subjects

75 CONDENSED MATTER PHYSICS
SUPERCONDUCTIVITY AND SUPERFLUIDITY
60 APPLIED LIFE SCIENCES
ALBUMINS
ATOMIC FORCE MICROSCOPY
BIOCHEMISTRY
CATTLE
CHANNELING
DIFFRACTION
FLUORESCENCE
FOURIER TRANSFORMATION
FUNCTIONS
INFRARED SPECTRA
INTERFACES
IRRADIATION
LAYERS
PHOTOCHEMISTRY
POLYSTYRENE
RADIATION EFFECTS
SPATIAL RESOLUTION
SPECTROSCOPY
SUBSTRATES
SURFACES