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Aims/Introduction: Severe diabetic macular edema (DME) is often resistant to anti‐vascular endothelial growth factor therapy. Steroids are particularly effective at reducing edema by suppressing inflammation; they are also used as an alternative to expensive anti‐vascular endothelial growth factor therapy in some patients. Therefore, the use of steroids in DME reflects an unmet need for anti‐vascular endothelial growth factor therapy. Notably, triamcinolone acetonide (TA) injections are widely used in Japan. Here, we evaluated the frequency of TA as an indicator of the efficacy of sodium–glucose cotransporter 2 inhibitors (SGLT2is) in DME treatment using a health insurance claims database. Materials and Methods: In this cohort study, we retrospectively analyzed the health insurance claims data of 11 million Japanese individuals from 2005 to 2019. The frequency and duration of TA injection after the initiation of SGLT2is or other antidiabetic drugs were analyzed. Results: Among the 2,412 matched patients with DME, the incidence rate of TA injection was 63.8 times per 1,000 person‐years in SGLT2i users and 94.9 times per 1,000 person‐years in non‐users. SGLT2is reduced the risk for the first (P = 0.0024, hazard ratio 0.66, 95% confidence interval 0.50–0.87), second (P = 0.0019, hazard ratio 0.53, 95% confidence interval 0.35–0.80) and third TA (P = 0.0053, hazard ratio 0.44, 95% confidence interval 0.25–0.80) injections. A subanalysis of each baseline characteristic of the patients showed that SGLT2is were effective regardless of the background factors. Conclusions: The use of SGLT2is reduced the frequency of TA injection in patients with DME. Therefore, SGLT2i therapy might be a novel, noninvasive and low‐cost adjunctive therapy for DME. [ABSTRACT FROM AUTHOR]

Studies have shown that COVID-19 school closures negatively impacted children’s well-being. We assessed the impact of school closures on children in Japan. We postulated that screen time and sleep habits during school closure would be associated with children’s mental health status.

A continuous downstream process of monoclonal antibody was developed based on the process characterization. Periodic‐counter current chromatography (PCCC) with two protein A columns was used for the capture step. For low pH virus inactivation (VI), a batch reactor was employed, which can work as a surge (buffer) tank. Flow‐through chromatography (FTC) with two connected columns of different separation modes (anion‐mixed mode and cation exchange) was designed as a polish step. After 24 h PCCC run, the collected pool was processed for VI. After adjusting pH and electric conductivity, the solution was fed to the two connected FTC columns for 24 h. Virus filter was also connected to the exit of the connected‐column. PCCC and FTC were run in parallel. Six runs of different feed rates (0.5–10 L/day) and feed concentrations (1–3.2 g/L) were performed with protein A columns of 1–5 mL and FTC columns of 3–10 mL. The largest run (feed rate 10 L/day, feed concentration 2 g/L) was carried out at a GMP facility with 15 mL protein A columns and 100 mL FTC columns. Good recovery and purity values were obtained for all runs. The process was found to be flexible and stable for feed fluctuations. Only three surge or pool tanks were needed in addition to the final product pool tank. [ABSTRACT FROM AUTHOR]

Aim: We assessed the effectiveness of sodium‐glucose co‐transporter 2 inhibitors (SGLT2is) in reducing the administration frequency of anti‐vascular endothelial growth factor (VEGF) agents in patients with diabetic macular oedema (DMO) using a health insurance claims database. Materials and Methods: This retrospective cohort study analysed health insurance claims data covering 11 million Japanese patients between 2005 and 2019. We analysed the frequency and duration of intravitreal injection of anti‐VEGF agents after initiating SGLT2is or other antidiabetic drugs. Results: Among 2412 matched patients with DMO, the incidence rates of anti‐VEGF agent injections were 230.1 per 1000 person‐year in SGLT2i users and 228.4 times per 1000 person‐year in non‐users, respectively, and the risk ratio for events was unchanged in both groups. Sub‐analysis of each baseline characteristic of the patients showed that SGLT2is were particularly effective in patients with a history of anti‐VEGF agent use [p =.027, hazard ratio (HR): 0.44, 95% confidence interval (CI): 0.22‐0.91]. SGLT2is reduced the risk for the first (p =.023, HR: 0.45, 95% CI: 0.22‐0.91) and second (p =.021, HR: 0.39, 95% CI: 0.17‐0.89) anti‐VEGF agent injections. Conclusions: There was no difference in the risk ratio for the addition of anti‐VEGF therapy between the two treatment groups. However, the use of SGLT2is reduced the frequency of anti‐VEGF agent administration in patients with DMO requiring anti‐VEGF therapy. Therefore, SGLT2i therapy may be a novel, non‐invasive, low‐cost adjunctive therapy for DMO requiring anti‐VEGF therapy. [ABSTRACT FROM AUTHOR]

Monoclonal antibody drug conjugate (ADCs) have received much attention as pharmaceutical agents for treating serious diseases such as cancer. However, it is difficult to separate them on the basis of the drug to antibody ratio, DAR. Hydrophobic chromatography (HIC) is commonly used for the analysis of the drug to antibody ratio, DAR. The retention of ADCs on HIC can be controlled by the hydrophobic nature of ADCs, depending on the mobile phase conditions. They are sometimes performed at the restricted conditions where the solubility is too low. Ion exchange chromatography (IEC) using electrostatic interaction is an orthogonal method to HIC. IEC is widely used because of its higher capacity than HIC. We investigated the retention behavior of the protein conjugated with surrogate drugs on IEC. The surrogate drugs employed are 7-diethylamino-3-(4’-maleimidylhenyl) 4-methylcoumarin (CPM), N-(1-pyrenyl) maleimide (NPM). Bovine serum albumin (BSA) was used as a model protein. The molar ratio (CPM and NPM to protein) was set to 3. The maleimide group of CPM and NPM reacts with the thiol group of the proteins. On the linear gradient elution experiments, the elution salt concentrations of the conjugated and non-conjugated proteins were measured to obtain chromatographic parameter of the number of binding sites, B.

Ion exchange chromatography is a widely used method for purification of all types of biomolecules in current biotechnological downstream processes. Knowledge on the binding behavior of proteins provides valuable insight for understanding the molecular mechanisms of protein interactions in a biological context. However, thermodynamic parameters such as enthalpy and entropy changes that characterize protein adsorption are still unknown. Isothermal titration calorimetry applications in biosciences has gained its merit to study binding of soluble molecules, protein inhibition, conformational changes, reaction kinetics, and protein adsorption. However, in the case of protein adsorption, a lot of complications arise since the usual models used to study protein interactions in solution are no longer valid. This work explains a detailed methodology for the obtention of adsorption enthalpy, entropy and Gibbs energy of protein adsorption, by using ITC together with equilibrium adsorption isotherms.

The antibody manufacturing process consists of the cell culture process using animal cells and the purification process including several chromatography steps and filtration steps. The purification process is important to obtain high quality bio-pharmaceutical products by removing impurities, and measurement of antibody concentrations is a fundamental and essential factor to control the manufacturing process and to confirm the yield of each purification step. Here we introduce the optical rotation method as a resolution for measuring antibody concentrations in the manufacturing process. Feasibility evaluation showed that optical rotation is applicable for measuring a wide range of antibody concentrations even in cell culture supernatants containing various contaminants. In addition, as examples of application to the process analytical technology, we show the possibility of the optical rotation method in the continuous cell culture and chromatography process, and the tangential flow filtration concentration process.

In this study, the swelling pressure of unsaturated, compacted bentonite-sand mixture was measured, which the all samples had experiment in dying-wetting history in suction changing. This test program used Kunigel V1 as sodium type. Constant volume swelling test was conducted out that swelling pressure was measured using a developed apparatus. A pressure sensor was mounted in the swelling test apparatus. Previously, the all samples deformed due to suction change (i.e. drying-wetting process in suction). Two types were prepared before applying suction that were initial unsaturated condition and saturated condition on same initial dry density. Swelling pressure and adsorbed water amount was monitored in the test, which lasted for one month.

There are few reports of the adoption of continuous processes in bioproduction, particularly the implementation of end‐to‐end continuous or integrated processes, due to difficulties such as feed adjustment and incorporating virus filtration. Here, we propose an end‐to‐end integrated continuous process for a monoclonal antibody (mAb) with three integrated process segments: upstream production processes with pool‐less direct connection, pooled low pH virus inactivation with pH control and a total flow‐through integrated polishing process in which two columns were directly connected with a virus filter. The pooled virus inactivation step defines the batch, and high impurities reduction and mAb recovery were achieved for batches conducted in succession. Viral clearance tests also confirmed robust virus reduction for the flow‐through two‐column chromatography and the virus filtration steps. Additionally, viral clearance tests with two different hollow fiber virus filters operated at flux ranging from 1.5 to 40 LMH (liters per effective surface area of filter in square meters per hour) confirmed robust virus reduction over these ranges. Complete clearance with virus logarithmic reduction value ≥4 was achieved even with a process pause at the lowest flux. The end‐to‐end integrated continuous process proposed in this study is amenable to production processes, and the investigated virus filters have excellent applicability to continuous processes conducted at constant flux. [ABSTRACT FROM AUTHOR]