Phage Display Antibody-Based Proteomic Device Using Resonance-Enhanced Detection
The combination of phage display antibody arrays with a novel nanotransducer technique based on resonant nanoparticles in a nanosandwiched film enables the sensitive parallel screening of proteins. Using
the resonance of nanoparticles with their induced mirror dipoles in a thin-film structure, limitations of fluorophores, such as unspecific background and nonvisibility to the eye, can be overcome, thereby
leading to an optical signal significantly more sensitive than that of standard colloid techniques. The signal can be both directly observed as a color change of a microdot at the sensor surface and tuned
throughout the visible range of the spectrum. Here we report the application of an optical chip using scFv-antibody-antigen interactions. Artificial scFv-antibodies against a variety of proteins, including
yeast enzymes and bovine serum albumin (as a standard), were constructed via Phage Display. These scFv-antibodies were then coated onto metal nanoclusters and bound to their antigens that were arrayed as
nanodroplets at the resonance layer of the chip. ScFv-Antibody-antigen interaction resulted in a visible array of microdots. Using resonance-enhanced absorption, the absorption signal of the spots was amplified
by one to two orders of magnitude (compared to colloid-based techniques). For quantitative analysis, either an 8-μm scanner or a CCD camera (resolution 4 μm) was employed to gain direct-reflection
spectra rather than unspecific scatter data (prone to dust and unspecific interaction). Our results demonstrate that this device enables high-throughput proteomics to overcome some limitations of fluorescence,
enzyme labels, and colloid techniques.
Keywords: METAL NANOPARTICLES; OPTICAL NANORESONANCE; PHAGE DISPLAY; PROTEIN BIOCHIP
Document Type: Research Article
Affiliations: 1: Kluyver Laboratory for Biotechnology, TU-Delft, 2628BC Delft, The Netherlands b Institut für Biochemie und Molekulare Zellbiologie, Universität Wien, Vienna, Austria 2: Institut für Biochemie und Molekulare Zellbiologie, Universität Wien, Vienna, Austria 3: Department of Biochemistry, University of Nijmegen, NL-6500 HB Nijmegen, The Netherlands 4: Kluyver Laboratory for Biotechnology, TU-Delft, 2628BC Delft, The Netherlands b Institut für Biochemie und Molekulare Zellbiologie, Universität Wien, Vienna, Austria
Publication date: 01 July 2002
- Journal for Nanoscience and Nanotechnology (JNN) is an international and multidisciplinary peer-reviewed journal with a wide-ranging coverage, consolidating research activities in all areas of nanoscience and nanotechnology into a single and unique reference source. JNN is the first cross-disciplinary journal to publish original full research articles, rapid communications of important new scientific and technological findings, timely state-of-the-art reviews with author's photo and short biography, and current research news encompassing the fundamental and applied research in all disciplines of science, engineering and medicine.
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