Surface plasmon resonance(SPR) systems are widely used for detailed characterization of antibody activities including antigen and Fc-receptor binding. During the later stages of development, where the focus is to ensu...Surface plasmon resonance(SPR) systems are widely used for detailed characterization of antibody activities including antigen and Fc-receptor binding. During the later stages of development, where the focus is to ensure that established critical quality attributes(CQAs) are maintained during cell culture,purification and formulation processes, analysis is simplified, and relative potencies are often determined. Here, simulation of binding data revealed that relative potency values, determined via parallel line analysis(PLA) and half maximal effective concentration(EC50) analysis accurately reflect changes in active concentration only if binding kinetics remain unchanged. Changes in the association rate constant shifted dose response curves, and therefore relative potencies, in the same way as changes in analyte concentration do. However, for interactions characterized by stable binding, changes in the dissociation rate constant did not result in any shift, suggesting that this type of change may go unnoticed in the dose response curve. Thus, EC50 and PLA analyses of dose response curves obtained with an anti-TNF-α antibody were complemented with the Biacore functionality for sensorgram comparison analysis, whereby changes in antigen and Fc-receptor binding profiles could be detected. Next, analysis of temperature stressed TNF-α antibody revealed that calibration free concentration analysis(CFCA) data correlated perfectly with relative potency values. Together, these results demonstrate that combinations of SPR based dose response curves, sensorgram comparison and CFCA can be used to strengthen the confidence in relative potency assessments, and suggest that SPR can potentially be used as a surrogate potency assay in the quality control of biotherapeutic medicines.展开更多
The tapered element oscillating microbalance with filter dynamics measurement system （TEOM-FDMS） is an instrument commonly employed by the French air quality monitoring network. This instrument is currently calibrat...The tapered element oscillating microbalance with filter dynamics measurement system （TEOM-FDMS） is an instrument commonly employed by the French air quality monitoring network. This instrument is currently calibrated with calibration weights traceable to SI but having value and mass differences between each of them that are not representative of real atmospheric particle mass measurements. Moreover, these calibration weights do not allow detection of any technical problems associated with either the TEOM-FDMS sampling system upstream of the mass measurement or the intrinsic TEOM-FDMS filtration system. Therefore, a calibration method was developed using a portable reference aerosol generator （PRAG） that produces known and stable particle mass concentrations over time. Here, we present the characterization of the PRAG system in terms of a reference range of particle masses between 30 - 10 and 3456 ± 83 μg at three sampling times. Its coupling with the TEOM-FDMS and a global comparison between the defined reference range of particle masses and the measured masses obtained with each TEOM-FDMS implicated in this study are also presented.展开更多
文摘Surface plasmon resonance(SPR) systems are widely used for detailed characterization of antibody activities including antigen and Fc-receptor binding. During the later stages of development, where the focus is to ensure that established critical quality attributes(CQAs) are maintained during cell culture,purification and formulation processes, analysis is simplified, and relative potencies are often determined. Here, simulation of binding data revealed that relative potency values, determined via parallel line analysis(PLA) and half maximal effective concentration(EC50) analysis accurately reflect changes in active concentration only if binding kinetics remain unchanged. Changes in the association rate constant shifted dose response curves, and therefore relative potencies, in the same way as changes in analyte concentration do. However, for interactions characterized by stable binding, changes in the dissociation rate constant did not result in any shift, suggesting that this type of change may go unnoticed in the dose response curve. Thus, EC50 and PLA analyses of dose response curves obtained with an anti-TNF-α antibody were complemented with the Biacore functionality for sensorgram comparison analysis, whereby changes in antigen and Fc-receptor binding profiles could be detected. Next, analysis of temperature stressed TNF-α antibody revealed that calibration free concentration analysis(CFCA) data correlated perfectly with relative potency values. Together, these results demonstrate that combinations of SPR based dose response curves, sensorgram comparison and CFCA can be used to strengthen the confidence in relative potency assessments, and suggest that SPR can potentially be used as a surrogate potency assay in the quality control of biotherapeutic medicines.
文摘The tapered element oscillating microbalance with filter dynamics measurement system （TEOM-FDMS） is an instrument commonly employed by the French air quality monitoring network. This instrument is currently calibrated with calibration weights traceable to SI but having value and mass differences between each of them that are not representative of real atmospheric particle mass measurements. Moreover, these calibration weights do not allow detection of any technical problems associated with either the TEOM-FDMS sampling system upstream of the mass measurement or the intrinsic TEOM-FDMS filtration system. Therefore, a calibration method was developed using a portable reference aerosol generator （PRAG） that produces known and stable particle mass concentrations over time. Here, we present the characterization of the PRAG system in terms of a reference range of particle masses between 30 - 10 and 3456 ± 83 μg at three sampling times. Its coupling with the TEOM-FDMS and a global comparison between the defined reference range of particle masses and the measured masses obtained with each TEOM-FDMS implicated in this study are also presented.
