Studying antibody-antigen interactions with fluorescence fluctuation spectroscopy.

Abstract:

:Antibodies are excellent binding proteins that have found numerous applications in biological research, biotechnology, and medicine. Characterization of their ligand binding properties has long been, and continues to be, the focus of many researchers. Antibodies are also perfect test systems which can be used for the evaluation of newly introduced biophysical techniques. Working with many different antibodies, we continuously implement the growing arsenal of methods offered by fluorescence fluctuation spectroscopy (FFS) and apply them for antibody research. In this chapter, we will describe applications of FFS for antibody binding characterization and also provide examples how studying of antibodies helps to develop and enhance the tool set offered by FFS technology. In addition to traditional affinity evaluations, we will describe how resolving molecular populations enables determinations of the binding stoichiometry and provides further information about the system. Even though all our examples include antibodies, the same experimental procedures can also serve well for characterizing various proteins and other ligand binding systems.

journal_name

Methods Enzymol

journal_title

Methods in enzymology

authors

Tetin SY,Ruan Q,Skinner JP

doi

10.1016/B978-0-12-405539-1.00005-1

subject

Has Abstract

pub_date

2013-01-01 00:00:00

pages

139-66

eissn

0076-6879

issn

1557-7988

pii

B978-0-12-405539-1.00005-1

journal_volume

519

pub_type

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