Evaluation of the impact of antibody fragments on aggregation of intact molecules via size exclusion chromatography coupled with native mass spectrometry.

Aggregates are recognized as one of the most critical product-related impurities in monoclonal antibody (mAb)-based therapeutics due to their negative impact on the stability and safety of the drugs. So far, investigational efforts have primarily focused on understanding the causes and effects of mAb self-aggregation, including both internal and external factors. In this study, we focused on understanding mAb stability in the presence of its monovalent fragment, formed through hinge cleavage and loss of one Fab unit (referred to as "Fab/c"), a commonly observed impurity during manufacturing and stability. The Fab/c fragments were generated using a limited IgdE digestion that specifically cleaves above the IgG1 mAb hinge region, followed by hydrophobic interaction chromatographic (HIC) enrichment. Two IgG1 mAbs containing different levels of Fab/c fragments were incubated under thermally accelerated conditions. A method based on size exclusion chromatography coupled with native mass spectrometry (SEC-UV-native MS) was developed and used to characterize the stability samples and identified the formation of heterogeneous dimers, including intact dimer, mAb-Fab/c dimer, Fab/c-Fab/c dimer, and mAb-Fab dimer. Quantitative analyses on the aggregation kinetics suggested that the impact of Fab/c fragment on the aggregation rate of individual dimer differs between a glycosylated mAb (mAb1) and a non-glycosylated mAb (mAb2). An additional study of deglycosylated mAb1 under 25°C accelerated stability conditions suggests no significant impact of the N-glycan on mAb1 total aggregation rate. This study also highlighted the power of SEC-UV-native MS method in the characterization of mAb samples with regard to separating, identifying, and quantifying mAb aggregates and fragments.

Innovative approach to waste management: utilizing stabilized municipal solid waste in road infrastructure.

In recent years, a sudden upsurge in the quantity of municipal solid waste (MSW) has been observed, and the status quo demands a constructive and economically viable solution. The use of stabilized municipal solid waste (SMSW) in road construction can help in reducing the burden on landfills and waste management authorities. In the existing study, SMSW was accumulated from the Okhla landfill which is situated in New Delhi that is rich in an organic content. This SMSW was then blended with soil (5%, 10%, and 15%) and bottom ash with varying percentages (10%, 20%, 30%) individually and a mix of soil and bottom ash in the ratio of 1:1 to reduce the content of organic matter. The blended sample was then tested to check its compaction value, California bearing ratio (CBR), unconfined compressive strength (UCS), durability, and scanning electron microstructure (SEM). The results indicated that the addition of bottom ash to SMSW decreases the maximum dry unit weight that varies between 1.65 and 1.51 KN/m3 while this value reduces to 1.72 to 1.67 KN/m3 in the case of the bottom ash-soil blend. Also, CBR value reduces to 25.50 to 18.00% in case of bottom ash and 25.89 to 21.92% for bottom ash-soil samples and inverse in the case of SMSW samples blended with soil ranges between 19.95 and 22.59%. The California bearing ratio value of all samples under soaked condition meets the minimum criteria (> 10%) as specified in IRC SP-72 for low-volume roads, but at the same time failed to meet the durability specifications. Thus, it is recommended to use this soil subgrade material in arid regions.

An Intercompany Perspective on Compatibility and In-use Stability Studies to Enable Administration Of Biopharmaceutical Drug Products.

In-use stability and compatibility studies are often used in biotherapeutic development to assess stability and compatibility of biologic drugs with diluents and/or administration components at relevant conditions for the target route of administration (commonly intravenous, subcutaneous or intramuscular), to assure that patient safety and product efficacy are maintained during clinical use. To gain an understanding of current industry approaches for in-use stability and compatibility studies, the Formulation Workstream of the BioPhorum Development Group (BPDG), an industry-wide consortium, conducted an inter-company collaboration exercise, which included five bench-marking surveys around in-use stability and compatibility studies of biologic drugs. The results of this industry collaboration provide insights into the practicalities of these studies and how they are being used to support administration of biologics from early clinical programs to marketed products. The surveys queried topics including regulatory strategies and feedback; clinical in-use formulation, patient and site considerations; clinical blinding, masking and placebo approaches; study setup, execution and reporting; and clinical in-use stability and compatibility testing to provide a comprehensive picture of the range of common industry practices. This paper discusses the survey results and presents various approaches which can be used to guide the strategy and design of an in-use stability and compatibility program based on clinical and biomolecule needs.

Role of Zingiber officinale and autochthonous probiotic Bacillus coagulans in feeds of Catla catla (Hamilton, 1822) for growth promotion, immunostimulation, histoprotection, and control of DNA damage.

Experiment was conducted to assess the impact of ginger (Zingiber officinale) as a dietary supplement with probiotic bacterium Bacillus coagulans on growth performance, hematological parameters, and non-specific immune parameters in Catla catla. An attempt was also made to investigate their effects on histology and extent of DNA damage against pathogenic bacterium Aeromonas hydrophila in C. catla. Ten dietary treatments were designed in triplicate in which five groups of fingerlings fed on diets containing different incorporation level of dried ginger at 0 g kg-1 (C1; basal diet 1 with duckweed as major protein source), 1 g kg-1 (G1), 5 g kg-1 (G2), 10 g kg-1 (G3), and 15 g kg-1 (G4) and other five groups of fingerlings fed on diets (G5-G9) containing different incorporation level of dried ginger at 0, 1, 5, 10, and 15 g kg-1 along with probiotic bacterium B. coagulans @ 3000 CFU ml-1 for 90 days. Results clearly revealed significantly (P < 0.05) high values of weight gain, growth % gain in body weight, specific growth rate (SGR), RBC and WBC count, phagocytic activity, respiratory burst activity, serum protein, and serum bactericidal activity in group of fingerlings fed on ginger supplemented diets which further improved with the inclusion of probiotic with best results in treatment G8. Histopathological study and COMET assay reflected that Z. officinale and probiotics protect extent of DNA damage and the digestive organs from the detrimental effects of pathogenic bacteria affirming their positive role and harmonious effects of probiotic bacterium with Z. officinale-supplemented diets.

An Industry Perspective on the Challenges of Using Closed System Transfer Devices with Biologics and Communication Guidance to Healthcare Professionals.

Closed system transfer devices (CSTDs) have been used with hazardous drugs for several decades. The goal of this whitepaper is to increase awareness among healthcare professionals, device manufacturers, regulators, and pharmaceutical/biotech companies on the potential issues around the use of CSTDs with biologic drug products to allow their informed use in clinics. Specifically, we discuss the key topics related to the use of CSTDs with biologics products, including components and materials of construction, a breakdown of regulatory, technical, clinical site-related risks and challenges associated with the use of CSTDs with biological products, gathered from stakeholder discussion at the IQ CSTD workshop, and considerations on current testing requirements and communication strategies to drive further dialog on the appropriate use of CSTDs. Given the technical challenges of using CSTDs with biologics, coupled with the current regulations surrounding CSTD approval and proper use, as well as a need for alignment and standardization to enable a consistent strategy for compatibility testing and communication of incompatibilities, it is recommended that global health authorities and other stakeholders seek to understand these issues, in order to alleviate these problems and keep healthcare workers and patients safe from harm.

A novel electronic skin patch for delivery and pharmacokinetic evaluation of donepezil following transdermal iontophoresis.

The nature of Alzheimer's disease limits the effectiveness of available oral treatments. The aim of this study was to assess the feasibility of transdermal iontophoretic delivery of donepezil in a hairless rat model as a potential treatment modality in Alzheimer's and to evaluate the effect of current densities on its pharmacokinetics. Donepezil loaded integrated Wearable Electronic Drug Delivery (WEDD(®)) patches supplied current levels of 0, 0.13, 0.26 and 0.39 mA. Plasma extracted donepezil was analyzed by HPLC. Noncompartmental analysis was used to characterize disposition of the drug. The amount delivered across hairless rat skin and areas under the curve (AUC) were found to rise in proportion to the current levels. Peak plasma levels of 0.094, 0.237 and 0.336 µg/ml were achieved at 0.13, 0.26 and 0.39 mA respectively. Time to peak plasma concentrations was after termination of current and same for all current levels. Transdermal elimination half-life was significantly increased from the true value of 3.2h due to depot formation, prolonging complete absorption of the drug. Donepezil was successfully delivered iontophoretically at levels sufficient to produce pharmacodymanic effect. Pharmacokinetic analysis demonstrated linear kinetics at the current levels used and flip flop kinetics following iontophoretic administration.