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

Citation

Wang, Hua; Castner, David G.; Ratner, Buddy D.; & Jiang, Shaoyi (2004). Probing the orientation of surface-immobilized immunoglobulin G by time-of-flight secondary ion mass spectrometry. Langmuir: the ACS journal of surfaces and colloids, 20(5), 1877-1887.

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.

Keyword(s)

Amino Acid Sequence
Animals
Antibodies, Monoclonal
Biosensing Techniques
Gonadotropins
Humans
Immunoglobulin Fab Fragments
Immunoglobulin Fc Fragments
Immunoglobulin G
Mice
Microscopy, Atomic Force
Sensitivity and Specificity
Spectrometry, Mass, Secondary Ion
Surface Properties

Reference Type

Journal Article

Secondary Title

Langmuir: the ACS journal of surfaces and colloids

Author(s)

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

Year Published

2004

Date Published

1078185600

Volume Number

Issue Number

Pages

1877-1887