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 Enzymoljournal_title
Methods in enzymologyauthors
Tetin SY,Ruan Q,Skinner JPdoi
10.1016/B978-0-12-405539-1.00005-1subject
Has Abstractpub_date
2013-01-01 00:00:00pages
139-66eissn
0076-6879issn
1557-7988pii
B978-0-12-405539-1.00005-1journal_volume
519pub_type
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