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  • Effect of antibody modifications on its biomolecular binding as determined by surface plasmon resonance.

Effect of antibody modifications on its biomolecular binding as determined by surface plasmon resonance.

Analytical biochemistry (2011-11-19)
Sandeep Kumar Vashist
ABSTRACT

A surface plasmon resonance (SPR)-based procedure was developed to determine the effect of antibody modifications on its biomolecular binding behavior. Mouse immunoglobulin G (IgG) was immobilized on a protein A-functionalized gold-coated SPR chip. Goat anti-mouse IgG and its various commercially available modifications (i.e., conjugated with atto 550, atto 647, tetramethylrhodamine isothiocyanate [TRITC], horseradish peroxidase [HRP], or biotin) were employed in exactly the same concentration for the detection of mouse IgG. The various modifications of goat anti-mouse IgG decreased its biomolecular binding to mouse IgG in the order of unmodified>HRP-labeled>atto 550-labeled>biotinylated>TRITC-labeled>atto 647-labeled.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Atto 550 Protein Labeling Kit, BioReagent, suitable for fluorescence
Sigma-Aldrich
Atto 550, for fluorescence, ≥90% (HPLC)
Sigma-Aldrich
Atto 647 NHS ester, BioReagent, suitable for fluorescence, ≥90% (HPLC)
Sigma-Aldrich
Atto 550 NHS ester, BioReagent, suitable for fluorescence
Sigma-Aldrich
Atto 647 maleimide, BioReagent, suitable for fluorescence, ≥90% (coupling to thiols)