title: Tailoring immobilization of immunoglobulin by excimer laser for biosensor applications.
abstract: The sheltered transfer and immobilization of rabbit anti-human antiserum immunoglobulin G (IgG) by matrix-assisted pulsed laser evaporation (MAPLE) are reported. The iced targets submitted to laser irradiation consisted of 0.2-2 mg/mL IgG blended or not with lipid (L-α-phosphatidylcholine dipalmitoyl) dissolved in distilled water-based saline buffer. Thin IgG coatings were obtained at room temperature onto glass, fused silica, or silicon substrates. Ten thousand subsequent laser pulses of 0.33, 0.5, or 0.67 J/cm(2) fluence were applied for the synthesis of each sample. Morphology and composition of the thin films were studied by optical, scanning, and atomic force microscopy and Fourier transformed infrared spectrometry. Optical labeling methods such as spectrofluorimetry and fluorescence microscopy were selected to verify the biosensor transduction principle because of their high sensitivity for detecting low amounts of antigen (IgG). Protein immobilization to the substrate surface was demonstrated for all obtained structures after immersion in the donkey anti-rabbit secondary antibody solution. The IgG transfer and immobilization onto substrates were improved by addition of lipid to MAPLE solutions.
authors.affiliation: National Institute for Lasers, Plasma and Radiation Physics, Magurele, Ilfov, Romania.
authors.forename: Felix
authors.initials: F
authors.lastname: Sima
authors.forename: Emanuel
authors.initials: E
authors.lastname: Axente
authors.forename: Carmen
authors.initials: C
authors.lastname: Ristoscu
authors.forename: Ion N
authors.initials: IN
authors.lastname: Mihailescu
authors.forename: Taras V
authors.initials: TV
authors.lastname: Kononenko
authors.forename: Ilya A
authors.initials: IA
authors.lastname: Nagovitsin
authors.forename: Galina
authors.initials: G
authors.lastname: Chudinova
authors.forename: Vitaly I
authors.initials: VI
authors.lastname: Konov
authors.forename: Marcela
authors.initials: M
authors.lastname: Socol
authors.forename: Ionut
authors.initials: I
authors.lastname: Enculescu
authors.forename: Livia E
authors.initials: LE
authors.lastname: Sima
authors.forename: Stefana M
authors.initials: SM
authors.lastname: Petrescu
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism*; D007074:Immunoglobulin G / Q000378:metabolism* / Q000648:ultrastructure; D054018:Lasers, Excimer; D008055:Lipids / Q000494:pharmacology; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D011817:Rabbits; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D055767:Immobilized Proteins; D007074:Immunoglobulin G; D008055:Lipids