Probing the orientation of surface-immobilized immunoglobulin G by time-of-flight secondary ion mass spectrometry

Hua Wang, David G. Castner, Buddy D. Ratner, Shaoyi Jiang

Research output: Contribution to journalArticle

134 Scopus citations

Abstract

Static time-of-flight secondary ion mass spectrometry (ToF-SIMS) is a powerful surface analysis technique for the characterization of protein films because of its chemical selectivity and surface sensitivity. In this study, static ToF-SIMS and principal component analysis (PCA), a multivariate data analysis method, were combined to probe the orientation of surface-immobilized immunoglobulin G (IgG). IgG orientation can enhance its ability to detect its antigen in immunoassay techniques. The IgG used in this work is the mouse monoclonal anti-human chorionic gonadotropin (anti-hCG). Anti-hCG films on different well-defined substrates have been studied using its F(ab′)2 and Fc fragments as references. Atomic force microscopy was used to characterize these protein films before static ToF-SIMS analysis. The results from PCA of ToF-SIMS spectra were related to the antibody primary amino acid composition and its three-dimensional structure.

Original languageEnglish (US)
Pages (from-to)1877-1887
Number of pages11
JournalLangmuir
Volume20
Issue number5
DOIs
StatePublished - Mar 2 2004

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ASJC Scopus subject areas

  • Materials Science(all)
  • Condensed Matter Physics
  • Surfaces and Interfaces
  • Spectroscopy
  • Electrochemistry

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