scholarly journals Forced Degradation Testing as Complementary Tool for Biosimilarity Assessment

2019 ◽  
Vol 6 (3) ◽  
pp. 62 ◽  
Author(s):  
Yan Felix Karl Dyck ◽  
Daniel Rehm ◽  
Jan Felix Joseph ◽  
Karsten Winkler ◽  
Volker Sandig ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Mass Spectrometry ◽  
Posttranslational Modification ◽  
Reference Product ◽  
Forced Degradation ◽  
High Similarity ◽  
Critical Quality Attributes ◽  
Forced Oxidation ◽  
Degradation Testing ◽  
Oxidation Susceptibility

Oxidation of monoclonal antibodies (mAbs) can impact their efficacy and may therefore represent critical quality attributes (CQA) that require evaluation. To complement classical CQA, bevacizumab and infliximab were subjected to oxidative stress by H2O2 for 24, 48, or 72 h to probe their oxidation susceptibility. For investigation, a middle-up approach was used utilizing liquid chromatography hyphenated with mass spectrometry (LC-QTOF-MS). In both mAbs, the Fc/2 subunit was completely oxidized. Additional oxidations were found in the light chain (LC) and in the Fd’ subunit of infliximab, but not in bevacizumab. By direct comparison of methionine positions, the oxidized residues in infliximab were assigned to M55 in LC and M18 in Fd’. The forced oxidation approach was further exploited for comparison of respective biosimilar products. Both for bevacizumab and infliximab, comparison of posttranslational modification profiles demonstrated high similarity of the unstressed reference product (RP) and the biosimilar (BS). However, for bevacizumab, comparison after forced oxidation revealed a higher susceptibility of the BS compared to the RP. It may thus be considered a useful tool for biopharmaceutical engineering, biosimilarity assessment, as well as for quality control of protein drugs.

scholarly journals PVP-H2O2 Complex as a New Stressor for the Accelerated Oxidation Study of Pharmaceutical Solids

Pharmaceutics ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 457 ◽  
Author(s):  
Dattatray Modhave ◽  
Brenda Barrios ◽  
Amrit Paudel
Keyword(s):  
Hydrogen Peroxide ◽  
Solid State ◽  
Pharmaceutical Products ◽  
Screening Methods ◽  
Forced Degradation ◽  
Pharmaceutical Solids ◽  
Drug Degradation ◽  
Oxidation Study ◽  
Degradation Reactions ◽  
Forced Oxidation

Reactive impurities, such as hydrogen peroxide in excipients, raise a great concern over the chemical stability of pharmaceutical products. Traditional screening methods of spiking impurities into solid drug-excipient mixtures oversimplify the micro-environment and the physical state of such impurities in real dosage form. This can lead to an inaccurate prediction of the long-term product stability. This study presents the feasibility of using a polyvinylpyrrolidone-hydrogen peroxide complex (PVP-H2O2) as an oxidative agent for the solid state forced degradation of a selected drug, vortioxetine HBr. The PVP-H2O2 complex was prepared and characterized using FT-IR spectroscopy. The tablet compacts were made using a mixture of solid PVP-H2O2 complex and crystalline vortioxetine HBr powder. The compacts were exposed to 40 °C/75% RH condition in open and closed states for different time intervals. The extent and the type of drug degradation were analysed using LC and LC-MS. The extent of degradation was higher in the samples stored at the open state as compared to the close state. The solution state forced oxidation was conducted to verify the peroxide induced degradation reactions. The results evidence the utility of the proposed solid-state stressor and the method for screening the sensitivity of drugs to the excipient reactive impurities involving peroxides in solid-state.

Oxidative stress-induced posttranslational modification of TRPV1 expressed in esophageal epithelial cells

2011 ◽  
Vol 301 (2) ◽  
pp. G230-G238 ◽  
Author(s):  
Etsuko Kishimoto ◽  
Yuji Naito ◽  
Osamu Handa ◽  
Hitomi Okada ◽  
Katsura Mizushima ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Posttranslational Modification ◽  
Transient Receptor Potential ◽  
Epithelial Cell Line ◽  
Transient Receptor Potential Vanilloid ◽  
Esophageal Mucosa ◽  
Chemokine Production ◽  
Modified Proteins ◽  
Esophageal Epithelial Cells

Human esophageal epithelium is continuously exposed to physical stimuli or to gastric acid that sometimes causes inflammation of the mucosa. Transient receptor potential vanilloid 1 (TRPV1) is a nociceptive, Ca2+-selective ion channel activated by capsaicin, heat, and protons. It has been reported that activation of TRPV1 expressed in esophageal mucosa is involved in gastroesophageal reflux disease (GERD) or in nonerosive GERD symptoms. In this study, we examined the expression and function of TRPV1 in the human esophageal epithelial cell line Het1A, focusing in particular on the role of oxidative stress. Interleukin-8 (IL-8) secreted by Het1A cells upon stimulation by capsaicin or acid with/without 4-hydroxy-2-nonenal (HNE) was measured by ELISA. Following capsaicin stimulation, the intracellular production of reactive oxygen species (ROS) was determined using a redox-sensitive fluorogenic probe, and ROS- and HNE-modified proteins were determined by Western blotting using biotinylated cysteine and anti-HNE antibody, respectively. HNE modification of TRPV1 proteins was further investigated by immunoprecipitation after treatment with synthetic HNE. Capsaicin and acid induced IL-8 production in Het1A cells, and this production was diminished by antagonists of TRPV1. Capsaicin also significantly increased the production of intracellular ROS and ROS- or HNE-modified proteins in Het1A cells. Moreover, IL-8 production in capsaicin-stimulated Het1A cells was enhanced by synthetic HNE treatment. Immunoprecipitation studies revealed that TRPV1 was modified by HNE in synthetic HNE-stimulated Het1A cells. We concluded that TRPV1 functions in chemokine production in esophageal epithelial cells, and this function may be regulated by ROS via posttranslational modification of TRPV1.

Cu isotope fractionation response to oxidative stress in a hepatic cell line studied using multi-collector ICP-mass spectrometry

2018 ◽  
Vol 410 (9) ◽  
pp. 2385-2394 ◽  
Author(s):  
María R. Flórez ◽  
Marta Costas-Rodríguez ◽  
Charlotte Grootaert ◽  
John Van Camp ◽  
Frank Vanhaecke
Keyword(s):  
Oxidative Stress ◽  
Mass Spectrometry ◽  
Cell Line ◽  
Isotope Fractionation ◽  
Hepatic Cell ◽  
Hepatic Cell Line ◽  
Icp Mass Spectrometry ◽  
Cu Isotope

scholarly journals A Novel and Potentially Multifaceted Dehydroascorbate Reductase Increasing the Antioxidant Systems is Induced by Beauvericin in Tomato

Antioxidants ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 435 ◽  
Author(s):  
Martina Loi ◽  
Silvana De Leonardis ◽  
Giuseppina Mulè ◽  
Antonio F. Logrieco ◽  
Costantino Paciolla
Keyword(s):  
Oxidative Stress ◽  
Mass Spectrometry ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Calvin Cycle ◽  
Mass Spectrometry Analysis ◽  
Antioxidant Systems ◽  
The Novel ◽  
Main Band ◽  
Chloroplast Mrna

Dehydroascorbate reductases (DHARs) are important enzymes that reconvert the dehydroascorbic acid (DHA) into ascorbic acid (ASC). They are involved in the plant response to oxidative stress, such as that induced by the mycotoxin beauvericin (BEA). Tomato plants were treated with 50 µM of BEA; the main antioxidant compounds and enzymes were evaluated. DHARs were analyzed in the presence of different electron donors by native and denaturing electrophoresis as well as by western blot and mass spectrometry to identify a novel induced protein with DHAR activity. Kinetic parameters for dehydroascorbate (DHA) and glutathione (GSH) were also determined. The novel DHAR was induced after BEA treatment. It was GSH-dependent and possessed lower affinity to DHA and GSH than the classical DHARs. Interestingly, the mass spectrometry analysis of the main band appearing on sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) revealed a chloroplast sedoheptulose 1,7-bisphosphatase, a key enzyme of the Calvin cycle, and a chloroplast mRNA-binding protein, suggesting that the DHA reducing capacity could be a side activity or the novel DHAR could be part of a protein complex. These results shed new light on the ascorbate-glutathione regulation network under oxidative stress and may represent a new way to increase the plant antioxidant defense system, plant nutraceutical value, and the health benefits of plant consumption.

scholarly journals Proteomic Analysis of Cerebrospinal Fluid Changes Related to Postmortem Interval

2006 ◽  
Vol 52 (10) ◽  
pp. 1906-1913 ◽  
Author(s):  
Erin J Finehout ◽  
Zsofia Franck ◽  
Norman Relkin ◽  
Kelvin H Lee
Keyword(s):  
Oxidative Stress ◽  
Mass Spectrometry ◽  
Cerebrospinal Fluid ◽  
Tandem Mass Spectrometry ◽  
Gel Electrophoresis ◽  
Protein Production ◽  
Postmortem Interval ◽  
Cytoskeletal Proteins ◽  
Tandem Mass ◽  
Concentration Change

Abstract Background: The study of proteins with altered production in postmortem cerebrospinal fluid (CSF) compared with antemortem CSF may improve the understanding of biochemical changes that occur immediately after death. Methods: Two CSF samples (1 antemortem and 1 postmortem) were collected from 7 patients and analyzed by 2-dimensional gel electrophoresis. An analysis was also performed to identify proteins that showed a correlation between concentration change and postmortem interval. Tandem mass spectrometry was used to identify the proteins. Results: Fifty-four protein spots were identified that showed a consistent and significant change in concentration in the postmortem CSF of all 7 patients (>3.5-fold, P <0.01). The proteins in these spots derive from a variety of functional groups, including cytoskeletal proteins, enzymes involved in glycolysis, and proteins that prevent oxidative stress. Fourteen protein spots were found to have an increase in production that correlated with postmortem interval. Conclusions: Changes in protein production of postmortem vs antemortem CSF were studied. The proteins observed to change production in the postmortem CSF include several proteins previously observed as potential stroke biomarkers.

Sign in / Sign up

Export Citation Format

Share Document