The photostability of the commonly used biotin-4-fluorescein probe

Richard A Haack; Kerry M Swift; Qiaoqiao Ruan; Richard J Himmelsbach; Sergey Y Tetin

doi:10.1016/j.ab.2017.05.019

The photostability of the commonly used biotin-4-fluorescein probe

Anal Biochem. 2017 Aug 15:531:78-82. doi: 10.1016/j.ab.2017.05.019. Epub 2017 May 22.

Authors

Richard A Haack¹, Kerry M Swift², Qiaoqiao Ruan², Richard J Himmelsbach², Sergey Y Tetin²

Affiliations

¹ Abbott Diagnostics Division, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL 60064-6016, USA. Electronic address: richard.haack@abbott.com.
² Abbott Diagnostics Division, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL 60064-6016, USA.

PMID: 28545867
DOI: 10.1016/j.ab.2017.05.019

Abstract

Biotin-4-fluorescein (B4F) is a commonly used fluorescent probe for studying biotin-(strept)avidin interactions. During a characterization study of an anti-biotin antibody, using B4F as the probe, we noticed a discrepancy in the expected and experimentally determined number of biotin binding sites. Analytical testing showed that the biotin moiety in the probe undergoes a photosensitized oxidation to produce a mixture of biotin sulfoxides which has the potential to impact the quantitation of binding sites using this fluorescent probe.

Keywords: Biotinylation; Fluorescein; Fluorophore; Photosensitizer; Strept(avidin); Sulfoxide.

MeSH terms

Biotin / analogs & derivatives*
Biotin / chemistry
Fluoresceins / chemistry*
Oxidation-Reduction
Photochemical Processes*
Streptavidin / chemistry

Substances

Fluoresceins
biotin-4-fluorescein
Biotin
Streptavidin