Hydrophobic interaction expanded bed adsorption chromatography (HI-EBAC) based facile purification of recombinant streptokinase from E. coli inclusion bodies. | LitMetric
The downstream processing of recombinant streptokinase (rSK), a protein used for dissolution of blood clots has been investigated employing Escherichia coli inclusion bodies obtained after direct chemical extraction followed by expanded bed adsorption chromatography (EBAC). Streptokinase was over-expressed using high cell density (final OD(600)=40) culture of recombinant E. coli, and an SK protein concentration of 1080 mg l(-1) was achieved. The wet cell pellet after centrifugation was re-suspended in 8M urea containing buffer resulting in direct extraction of almost 97% of cellular proteins into solution. Compared to mechanical disruption using sonication, the direct extraction helped in simultaneous cell lysis and inclusion body (IB) solubilization in a single integrated step. The post-extraction solution containing cell debris and cellular proteins was diluted and directly loaded on to an EBAC column containing Streamline phenyl, without clarification. By passing the solution four times through the column and using 1M NaCl during loading, 82.7% rSK activity could be recovered in the 10mM sodium phosphate buffer used for elution. A 3-fold increase in specific activity of rSK, from 0.18 x 10(5) in cell lysate to 0.53 x 10(5)IU mg(-1) resulted after this step. rSK was further purified to near-homogeneity (specific activity=0.96 x 10(5)IU mg(-1)) by a subsequent ion-exchange step operated in packed bed mode. An overall downstream recovery of 63% rSK was achieved after EBAC and ion exchange chromatography. The paper thus describes the purification of rSK using a three-step regime involving simple, efficient and highly facile steps.
Quantifying the mechanical properties of coronary arterial walls could provide meaningful information for the diagnosis, management, and treatment of coronary artery diseases. Since patient-specific coronary samples are not available for patients requiring continuous monitoring, direct experimental testing of vessel material properties becomes impossible. Current coronary models typically use material parameters from available literature, leading to significant mechanical stress/strain calculation errors.
With the exacerbation of population aging in China, policies have been formulated to provide elderly medical care resources. However, research on the matching situation of these resources with the elderly population (EP) is relatively scarce. This study collected data on elderly medical care resources and the EP from various provinces in mainland China from 2011 to 2017.
The common seahorse, Hippocampus kuda, is widely distributed throughout the Indo-Pacific region, occurring from East Africa to Hawaii, where it typically inhabits diverse ecosystems such as estuaries, lagoons, coastal zones, and seagrass beds. Despite its extensive geographic range, reports of its occurrence in the Red Sea remains limited, primarily restricted to the northern region of the Red Sea. Here, we report a new distributional record of H.
Introduction: Previous studies have shown that patients in the emergency department (ED) are frequently given incomplete discharge instructions that are written at least four grade levels above the recommended sixth-grade reading level, leading to poor understanding. Our aims in this study were to implement standardized discharge instructions containing six key components written at a more appropriate reading level for common emergency department (ED) diagnoses to improve patient understanding.
Methods: We conducted this study in a 20-bed ED at an urban Veteran's Administration hospital.
Does Cannabis Legalization Endanger Children and Adolescents? The Cannabis Act (CanG) came into force on April 1, 2024. It regulates the "controlled supply of cannabis to adults for recreational purposes". In the USA, legalization has contributed to a decrease in the risk perception of the health hazards of consumption and an increase in risky consumption patterns, including among adolescents.
