Enhancement of microsphere specificity to purify human serum albumin from blood plasma.

Efforts to rapidly and easily supply purified human serum albumin (HSA) in remote areas during a pandemic are challenging. Here, we developed an HSA-imprinted microsphere (HSAIM) as a matrix to purify HSA from blood plasma on a small scale. HSAIMs were generated by encapsulating silica-3-(2-imidazoline-1-yl) propyltriethoxysilane-Cu2+-HSA (SiO2-IMEO-Cu2+-HSA) into agarose gel, and the stereospecific template for HSA was obtained by eluting the agarose gel. The physicochemical properties and performance of HSAIM were evaluated, and HSAIMs were applied to purify HSA from human blood plasma. Spherical HSAIMs had an average diameter of 51.2 ± 6.1 µm. HSAIMs had a maximum adsorption of 8.77 × 10-2 µmol HSA g-1 with an imprinting factor of 2.83, and the selectivity factors of BSA, thrombin and IgG were 0.96, 0.59 and 0.26, respectively. HSAIMs had a dynamic binding capacity (DBC10%) of 5.94 × 10-2 µmol HSA g-1 and could be reused up to 10 cycles with an ultimate recovery of 55.92%. HSA adsorption kinetics of HSAIM fitted to a pseudo-second-order mechanism, and HSA binding characteristics fit with a Sips isotherm model. For practical purposes, an initial blood plasma sample containing 24.9 ± 2.5 mg protein was pretreated with ethanol yielding 14.5 ± 4.6 mg protein, and further purification with HSAIM yielded 3.6 ± 1.1 mg protein. Starting with a blood plasma sample containing 149 type proteins, a single protein identified as HSA was obtained after final purification step with the HSAIM column, indicating that HSAIMs stereospecifically bound HSA. Hence, HSAIM was promising for blood plasma purification on a small scale.

Molecular modeling of Ruellia tuberosa L compounds as a-amylase inhibitor: an in silico comparation between human and rat enzyme model.

Inhibition of α-amylase is an important strategy to control post-prandial hyperglycemia. The present study on Ruellia tuberosa, known as traditional anti-diabetic agent, is being provided in silico study to identify compounds inhibiting α-amylase in rat and human. Compounds were explored from PubChem database. Molecular docking was studied using the autodock4. The interactions were further visualized and analyzed using the Accelrys Discovery Studio version 3.5. Binding energy of compounds to α-amylase was varying between -1.92 to -6.66 kcal/mol in rat pancreatic alpha amylase and -3.06 to -8.42kcal/mol in human pancreatic alpha amylase, and inhibition konstanta (ki) was varying between 13.12- 39460µM in rat and 0.67-5600µM in human. The docking results verify that betulin is the most potential inhibitor of all towards rat model alpha amylase and human alpha amylase. Further analysis reveals that betulin could be a potential inhibitor with non-competitive pattern like betulinic acid. In comparison, betulin has smaller Ki (0.67µM) than acarbose (2.6 µM), which suggesting that betulin is more potential as inhibitor than acarbose, but this assumption must be verified in vitro.