This project strives to synthesize a novel dye/chitosan complex that will readily purify lactate dehydrogenase (LDH) from a homogenized chicken breast solution via affinity column chromatography. In doing so, a fiscally efficient alternative to Cibacron Blue 3-GA (CB3), a highly effective dye complex, will be elucidated. This project arose due to CB3’s price and absence from the market. Choosing an appropriate dye required the use of a molecular docking server to calculate the free binding energies (FBE) of various dyes to LDH. It was concluded that Eriochrome Black T (EBT) would most efficiently bind to LDH’s receptor site. This was surmised by comparing the EBT’s FBE of -7.83 kcal/mol to CB3’s FBE of -9.01 kcal/mol. It should also be noted that Calmagite is a worthy substitute to CB3, given its FBE of -6.67 kcal/mol. Following these calculations, a synthetic plan will be devised. The functional groups of interest are the sulfonate and sulfonic acid on EBT and Calmagite respectively. These will lend themselves to creating microwave-assisted sulfonamide linkages to chitosan. Ultimately, the successfully synthesized product(s) will be used in lieu of CB3 to provide a more affordable method to purify LDH.
Poster, Final Report Form, Research Report