ABSTRACT
Early-stage antibody discovery and engineering typically require the cloning, expression, and screening of large numbers of proteins. Normally, DNA fragments encoding proteins of interest are cloned into extra-chromosomal plasmids that are amplified in Escherichia coli. Following purification from the bacteria, the plasmids are introduced into appropriate cells, and the expressed recombinant proteins screened for desired binding or function in a high-throughput manner. Even in a 96-well plate format, plasmid purification from E. coli is typically a labor intensive and time-consuming process. To further accelerate our existing biotherapeutic discovery workflows we designed, qualified, and enabled a fully integrated high-throughput plasmid purification and quantification workstation which we have termed AMPS (Automated Miniprep Plasmid Station). Using components from a commercially available kit, AMPS can purify plasmid preparations from twenty 96-deep-well plates of E. coli cultures, measure DNA absorbance at 260 nm, calculate plasmid concentrations, and prepare 96-deep-well plates for mammalian expression in an operator-independent manner. Plasmid yields and concentrations are equivalent to those obtained off-line. Furthermore, the quality of the DNA purified on the AMPS is equivalent to that obtained off-line in terms of DNA topology, and absence of contaminating bacterial chromosomal DNA and RNA. Most importantly, plasmids purified on the AMPS provide similar antibody titers following transfection in CHO cells as plasmids purified off-line. The AMPS bridges high-throughput E. coli colony picking capabilities typically available in an automation lab with downstream CHO expression needs and will facilitate screening of large numbers of biotherapeutics in binding and cell assay screens.
Subject(s)
DNA , Escherichia coli , Animals , Cricetinae , Cricetulus , DNA/genetics , DNA, Bacterial , Escherichia coli/genetics , Plasmids/genetics , Recombinant Proteins/geneticsABSTRACT
Social vulnerability has proved to be an independent risk factor for hypoglycemia in patients with diabetes. In some countries, patients who are in a vulnerable situation are assisted in the public health system which provides free medical care. This study compares the prevalence of hypoglycemia among patients with type 2 diabetes (T2D), in public versus private sector and its relationship with social vulnerability. This multicentric descriptive study included 600 patients with T2D from public and private care institutions of Argentina. Socioeconomic level (SEL) was evaluated through the Marketing Argentinean Association survey. Number of severe, documented symptomatic and asymptomatic hypoglycemias were registered. Among the patients included, 66% were assisted in the public sector. The 41% of patients (n = 246) registered at least 1 episode of any hypoglycemia event being more prevalent in the public sector compared to the private sector (50% vs. 22%). In the adjusted analysis it was observed a greater risk of hypoglycemia in public sector (OR 4, 95% CI 2.65-6.04) and in patients that did not have diabetological education (OR 2.28 95% CI 1.35-3.84). Similarly, unemployment (OR 5.04 95% CI 2.69-9.46), and marginal SEL (OR 60.79 95% CI 14.89-248.13) increased the risk of hypoglycemia. Several factors related to social vulnerability as unemployment, marginal SEL and poor sanitary education showed a significant increase in the hypoglycemia risk. Professionals working with people with diabetes must take into account these factors for a safe treatment of the disease.
La vulnerabilidad social ha demostrado ser un factor de riesgo independiente de hipoglucemia en pacientes con diabetes. Los pacientes que se encuentran en situación de vulnerabilidad social reciben asistencia en el sistema de salud pública que brinda atención médica gratuita. Este estudio compara la prevalencia de hipoglucemia en pacientes con diabetes tipo 2 en el sector público frente al privado y su relación con la vulnerabilidad social. Se realizó un estudio multicéntrico descriptivo que incluyó 600 pacientes con diabetes tipo 2 de instituciones de atención pública y privada de Córdoba. El nivel socioeconómico se evaluó a través de la encuesta de la Asociación Argentina de Marketing que evalúa la dimensión social, educativa y económica para estratificar el nivel socioeconómico. Se registró el número de hipoglucemias graves, documentadas sintomáticas y asintomáticas. El 66% de los pacientes pertenecían al sector público. El 41% de los pacientes (n = 246) registró al menos 1 episodio de cualquier evento de hipoglucemia. En el análisis ajustado, se observó un mayor riesgo de hipoglucemia en el sector público (OR 4, 95% CI 2.65-6.04), en pacientes que no tenían educación diabetológica previa (OR 2.28, 95% CI 1.35-3.84), en desempleados (OR 5.04, 95% CI 2.69-9.46) y en aquellos con nivel socioeconómico marginal (OR 60.79 95% CI 14.89-248.13). Factores relacionados con la vulnerabilidad social como el desempleo, el nivel socioeconómico marginal y educación sanitaria deficiente mostraron un aumento en el riesgo de hipoglucemia. Los profesionales que trabajan con personas con diabetes deben tener en cuenta estos factores para un tratamiento seguro de la enfermedad.
Subject(s)
Humans , Male , Female , Middle Aged , Aged , Hospitals, Private/statistics & numerical data , Delivery of Health Care/statistics & numerical data , Diabetes Mellitus, Type 2/epidemiology , Hospitals, Public/statistics & numerical data , Hypoglycemia/epidemiology , Argentina/epidemiology , Socioeconomic Factors , Logistic Models , Prevalence , Cross-Sectional Studies , Risk Factors , Public Sector , Private Sector , Vulnerable Populations/statistics & numerical data , Diabetes Complications/epidemiology , Diabetes Mellitus, Type 2/physiopathology , Social Determinants of Health , Hypoglycemia/physiopathologyABSTRACT
Social vulnerability has proved to be an independent risk factor for hypoglycemia in patients with diabetes. In some countries, patients who are in a vulnerable situation are assisted in the public health system which provides free medical care. This study compares the prevalence of hypoglycemia among patients with type 2 diabetes (T2D), in public versus private sector and its relationship with social vulnerability. This multicentric descriptive study included 600 patients with T2D from public and private care institutions of Argentina. Socioeconomic level (SEL) was evaluated through the Marketing Argentinean Association survey. Number of severe, documented symptomatic and asymptomatic hypoglycemias were registered. Among the patients included, 66% were assisted in the public sector. The 41% of patients (n = 246) registered at least 1 episode of any hypoglycemia event being more prevalent in the public sector compared to the private sector (50% vs. 22%). In the adjusted analysis it was observed a greater risk of hypoglycemia in public sector (OR 4, 95% CI 2.65-6.04) and in patients that did not have diabetological education (OR 2.28 95% CI 1.35-3.84). Similarly, unemployment (OR 5.04 95% CI 2.69-9.46), and marginal SEL (OR 60.79 95% CI 14.89-248.13) increased the risk of hypoglycemia. Several factors related to social vulnerability as unemployment, marginal SEL and poor sanitary education showed a significant increase in the hypoglycemia risk. Professionals working with people with diabetes must take into account these factors for a safe treatment of the disease.
La vulnerabilidad social ha demostrado ser un factor de riesgo independiente de hipoglucemia en pacientes con diabetes. Los pacientes que se encuentran en situación de vulnerabilidad social reciben asistencia en el sistema de salud pública que brinda atención médica gratuita. Este estudio compara la prevalencia de hipoglucemia en pacientes con diabetes tipo 2 en el sector público frente al privado y su relación con la vulnerabilidad social. Se realizó un estudio multicéntrico descriptivo que incluyó 600 pacientes con diabetes tipo 2 de instituciones de atención pública y privada de Córdoba. El nivel socioeconómico se evaluó a través de la encuesta de la Asociación Argentina de Marketing que evalúa la dimensión social, educativa y económica para estratificar el nivel socioeconómico. Se registró el número de hipoglucemias graves, documentadas sintomáticas y asintomáticas. El 66% de los pacientes pertenecían al sector público. El 41% de los pacientes (n = 246) registró al menos 1 episodio de cualquier evento de hipoglucemia. En el análisis ajustado, se observó un mayor riesgo de hipoglucemia en el sector público (OR 4, 95% CI 2.65-6.04), en pacientes que no tenían educación diabetológica previa (OR 2.28, 95% CI 1.35-3.84), en desempleados (OR 5.04, 95% CI 2.69-9.46) y en aquellos con nivel socioeconómico marginal (OR 60.79 95% CI 14.89-248.13). Factores relacionados con la vulnerabilidad social como el desempleo, el nivel socioeconómico marginal y educación sanitaria deficiente mostraron un aumento en el riesgo de hipoglucemia. Los profesionales que trabajan con personas con diabetes deben tener en cuenta estos factores para un tratamiento seguro de la enfermedad.
