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Vertical Scanning Interferometry for Label-Free Detection of Peptide-Antibody Interactions

Palermo, Andrea; Thelen, Richard; Weber, Laura; Foertsch, Tobias; Rentschler, Simone; Hackert, Verena; Syurik, Julia; Nesterov-Mueller, Alexander

Abstract:
Peptide microarrays are a fast-developing field enabling the mapping of linear epitopes in the immune response to vaccinations or diseases and high throughput studying of protein-protein interactions. In this respect, a rapid label-free measurement of protein layer topographies in the array format is of great interest but is also a great challenge due to the extremely low aspect ratios of the peptide spots. We have demonstrated the potential of vertical scanning interferometry (VSI) for a detailed morphological analysis of peptide arrays and binding antibodies. The VSI technique is shown to scan an array area of 5.1 square millimeters within 3–4 min at a resolution of 1.4 μm lateral and 0.1 nm vertical in the full automation mode. Topographies obtained by VSI do match the one obtained by AFM measurements, demonstrating the accuracy of the technique. A detailed topology of peptide-antibody layers on single spots was measured. Two different measurement regions are distinguished according to the antibody concentration. In the case of weakly diluted serum, the thickness of the antibody layer is independent of the serum dilution and corresponds to the physical thickness of the accumulated antibody layer. ... mehr

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Verlagsausgabe §
DOI: 10.5445/IR/1000093167
Veröffentlicht am 02.04.2019
Coverbild
Zugehörige Institution(en) am KIT Institut für Mikrostrukturtechnik (IMT)
Publikationstyp Zeitschriftenaufsatz
Jahr 2019
Sprache Englisch
Identifikator ISSN: 2571-5135
urn:nbn:de:swb:90-931670
KITopen-ID: 1000093167
Erschienen in High-Throughput
Band 8
Heft 2
Seiten Article: 7
Bemerkung zur Veröffentlichung Gefördert durch den KIT-Publikationsfonds
Vorab online veröffentlicht am 27.03.2019
Schlagworte peptide array; vertical scanning interferometry; atomic force microscopy; serum antibodies
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