Affinity chromatography has been used in the production of the enzyme cofactor biotin labelled with the 35S isotope.
Researchers at the Johns Hopkins Bloomberg School of Public Health in Baltimore explain in BMC Biotechnology that isotopes of sulphur, hydrogen and carbon have been used in the past to track the enzyme cofactor's metabolic origins, as well as how it attaches to proteins.
However, labelling it with the 35S isotope has resulted in predominantly oxidised biotin species in some cases.
Other problems have included low specific activity of the produced biotin, along with a generally low yield.
Using affinity chromatography to purify biotinylated Plasmosium falciparum (PfBCCP-79), followed by acid hydrolysis to liberate free biotin, the scientists were able to demonstrate the efficacy of the process.
The purified PfBCCP-79 was obtained with high yield, expressed at high levels and seen to be soluble.
BMC Biotechnology provides open-access research into manipulating biological entities in order to use them in experiments.
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