Biosimilar drug products must have a demonstrated similarity with respect to the reference product’s molecules in order to ensure both the effectiveness of the drug and the patients’ safety. In the paper the fusion framework of a highly sensitive NMR fingerprinting approach for conformational changes and mathematically-based biosimilarity metrics was introduced. The final goal was to translate the complex spectral information into biosimilarity scores, which were then used to estimate the degree of similarity between the biosimilar and the reference product. The proposed method was successfully applied to a small protein, i.e., filgrastim (neutropenia treatment), and a relatively large protein, i.e., monoclonal antibody rituximab (lymphoma treatment).
COBISS.SI-ID: 5986330
The purpose of this work was characterization of monoclonal antibody aggregation process and identification of stability factors that could be used as indicators of aggregation propensity. Conformational stability (melting temperature), colloidal stability (dynamic interaction parameter) and aggregation kinetics were assessed for two different IgG monoclonal antibody subclasses. Aggregation was induced by exposing the mAbs samples in different formulations at concentrations from 1 mg/mL to 50 mg/mL to elevated temperature. The correlation between aggregation kinetics and colloidal and conformational stability factors was evaluated and dynamic interaction parameter was found to be a promising predictor of aggregation propensity of monoclonal antibodies. This work is a contribution towards prediction of aggregation propensity and mechanisms at high protein concentration as a part of a high throughput, low resource screening method applicable to actual systems used for development and production of biopharmaceuticals.
COBISS.SI-ID: 3097956
In this paper Interferon-alpha (IFN) was conjugated via its N-terminal amino group by reductive amination to ?-aldehyde functional comb-shaped PolyPEG polymers (50 and70 kDa) and to linear PEG (30 kDa). Both PolyPEG-IFN conjugates retained a similar potency as that of the marketed comparator (PEGASYS®), whereas the linear PEG-IFN conjugate potency was greater. All conjugates showed extended half-lives compared to that of naked IFN, with the PolyPEG conjugates exhibiting the longest half-lives and the linear PEG conjugate, the shortest. Viscosity analysis showed that the linear PEG-IFN conjugate was over twice as viscous as both PolyPEG conjugates. Taken together, this work demonstrates the potential of PolyPEG conjugation to therapeutic proteins as a novel tool for optimizing pharmacokinetic profiles in a way that potentially allows administration of high-dose formulations because of lower conjugate viscosity.
COBISS.SI-ID: 1586268
In this paper the effects of seven different chromatographic parameters and five sample preparation parameters in a high performance liquid chromatography (HPLC) method for assay determination of benzalkonium chloride (BKC) in a nasal formulation were evaluated using two fractional factorial experimental designs. The design space of the analytical method was modeled using Umetrics Modde software and the optimal method conditions were predicted. The optimal method was validated for linearity, accuracy and precision. The use of experimental designs enables obtaining the maximum amount of information with the least possible number of experiments. Such designs are an economical manner in evaluating a variety of different factors and their interactions
COBISS.SI-ID: 1579612
In this study we have characterized the cell line RPMI 2650 and evaluated different culture conditions for an in vitro model for nasal mucosa. Cells were cultured in media MEM at the air–liquid (A–L) or liquid–liquid (L–L) interface for one or three weeks. Different cryopreservation methods and cell culture techniques were evaluated with immunolabelling of junctional proteins, ultrastructural analysis using electron microscopy, transepithelial electrical resistance (TEER) measurements, permeation studies with dextran and jacalin, and gene expression profiling of 84 drug transporters. Cells grown at the A-L interface formed more layers and exhibited a higher TEER and lower dextran and jacalin permeability than at the L-L interface, where cells morphologically exhibited a more differentiated phenotype. The RPMI 2650 cells form a polarized epithelium resembling nasal mucosa. However, different culture conditions have a significant effect on cell ultrastructure, barrier integrity, and gene expression, and should be considered when using this cell line as an in vitro model for drug permeability studies and screening of nasal drug candidates.
COBISS.SI-ID: 1567580