Date of this Version
Ford, A. (2019). Approaches to Synthesis of Biotin-Thiol Conjugates for Affinity Purification. Undergraduate Honors Thesis. University of Nebraska-Lincoln.
The interaction between biotin (vitamin H) and the protein, streptavidin, is one of the strongest known interactions between a protein and its substrate, which can allow for even trace quantities of desired proteins to be separated from a mixture of molecules when this interaction is exploited. The purpose of this research was to synthesize a novel biotin derivative, which includes a thiol group appended on a hydrophilic linker. Through linkage with the thiol functionality of the biotin derivative, the synthesized product was then intended to be applied to the formation of conjugates with drugs and other molecules thought to have strong interactions with unknown proteins. The strength and specificity of the biotin-streptavidin interactions could then be utilized for affinity purification of the unknown proteins. Herein, we provide promising results for successful synthesis of the finalized biotin derivative, which will be involved in the bioconjugation. This linked product can then be selectively purified and detected through the exploitation of the high affinity between biotin and streptavidin. This interaction is much more effective than trying to isolate drug-protein interactions independently where the attraction is much weaker. Thus, application of this interaction will aid in the purification of a target with extreme accuracy in order to isolate the desired drug after its interaction with proteins.