Ultrafiltration as an alternative purification method for bovine serum albumin-fluorescein isothiocyanate bioconjugate

TL;DR

This paper explores using ultrafiltration to purify a fluorescently tagged protein, comparing it to commercial methods and testing its effectiveness in lab experiments.

Contribution

The study introduces ultrafiltration as a novel purification method for FITC-labeled BSA bioconjugates.

Findings

Ultrafiltration successfully retained the conjugated protein while removing unreacted FITC.

The purified product performed comparably to a commercial product in UV-Vis spectroscopy.

The method was effective for in vitro bioassay applications.

Abstract

The bioconjugation of proteins with fluorescent probes is a popular method for protein visualization and has widespread applications across scientific fields. This study assessed ultrafiltration as an alternative purification method following the bioconjugation reaction that tags fluorescein isothiocyanate (FITC) to bovine serum albumin (BSA). Success of the purification was determined by (1) UV-Vis spectroscopy, comparing our sample to a commercially available product, and (2) use in an in vitro bioassay. Centrifugal ultrafiltration devices with a molecular weight cut off below the molecular weight of the conjugated protein were successful in retaining the purified product and clearing unreacted FITC from solution.

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