Detection of Beta-Glucan Contamination in Nanotechnology-Based Formulations

Neun, Barry; Cedrone, Edward; Potter, Timothy; Crist, Rachael M.; Dobrovolskaia, Marina A.

doi:10.3390/molecules25153367

Cited by 17 publications

(34 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nevertheless, the activation of all pathways results in the cleavage of the C3 component and the release of C3a, which is thus utilized to monitor for complement activation. Values are considered relevant when released C3a are more than 2-fold higher than control assay [ 35 ]. As shown in Figure 4 D for the tested NP, the C3a release was always ≤1.4-fold higher than the control.…”

Section: Resultsmentioning

confidence: 99%

Systematic Modification and Evaluation of Enzyme-Loaded Chitosan Nanoparticles

Lino

Leandro

Figueiredo

et al. 2021

IJMS

View full text Add to dashboard Cite

Polymeric-based nano drug delivery systems have been widely exploited to overcome protein instability during formulation. Presently, a diverse range of polymeric agents can be used, among which polysaccharides, such as chitosan (CS), hyaluronic acid (HA) and cyclodextrins (CDs), are included. Due to its unique biological and physicochemical properties, CS is one of the most used polysaccharides for development of protein delivery systems. However, CS has been described as potentially immunogenic. By envisaging a biosafe cytocompatible and haemocompatible profile, this paper reports the systematic development of a delivery system based on CS and derived with HA and CDs to nanoencapsulate the model human phenylalanine hydroxylase (hPAH) through ionotropic gelation with tripolyphosphate (TPP), while maintaining protein stability and enzyme activity. By merging the combined set of biopolymers, we were able to effectively entrap hPAH within CS nanoparticles with improvements in hPAH stability and the maintenance of functional activity, while simultaneously achieving strict control of the formulation process. Detailed characterization of the developed nanoparticulate systems showed that the lead formulations were internalized by hepatocytes (HepG2 cell line), did not reveal cell toxicity and presented a safe haemocompatible profile.

show abstract

Section: Resultsmentioning

confidence: 99%

Systematic Modification and Evaluation of Enzyme-Loaded Chitosan Nanoparticles

Lino

Leandro

Figueiredo

et al. 2021

IJMS

View full text Add to dashboard Cite

show abstract

“…As an improvement, the Limulus amebocyte lysate (LAL) test detects the hemolymph coagulation of the American horseshoe crab Limulus polyphemus when in the presence of bacterial endotoxin/LPS and is used as a standard for bacterial contamination [ 86 , 87 ]. However, this assay is specific for endotoxin, not general pyrogens [ 31 ], and has reduced specificity in the presence of fungal β-glucans because the horseshoe crab lysate used for this assay contains two proteins that trigger activation of the proteolytic cascade: factor C is specific to the presence of endotoxin while factor G is specific to β-glucans [ 88 , 89 ]. Knowing this, a modified version of the LAL assay containing glucan-blocking reagents or recombinant factor C overcomes β-glucan interference during endotoxin detection [ 90 ].…”

Section: Methods For Iimi Detectionmentioning

confidence: 99%

“…While β-(1,3)-d-glucans are not as immunologically potent as bacterial endotoxins, requiring μg/mL concentrations as compared to the endotoxin pg/mL concentrations to elicit an immunomodulatory response, they are a common IIMI present in many pharmaceutical products and solutions [ 89 ]. Moreover, while there is currently no compendial standard for β-glucan detection or acceptable levels, a modified version of the LAL assay is growing in popularity [ 90 ].…”

Section: Methods For Iimi Detectionmentioning

confidence: 99%

Innate Immunity Modulating Impurities and the Immunotoxicity of Nanobiotechnology-Based Drug Products

Holley

Dobrovolskaia

2021

Molecules

Self Cite

View full text Add to dashboard Cite

Innate immunity can be triggered by the presence of microbial antigens and other contaminants inadvertently introduced during the manufacture and purification of bionanopharmaceutical products. Activation of these innate immune responses, including cytokine secretion, complement, and immune cell activation, can result in unexpected and undesirable host immune responses. These innate modulators can also potentially stimulate the activation of adaptive immune responses, including the formation of anti-drug antibodies which can impact drug effectiveness. To prevent induction of these adverse responses, it is important to detect and quantify levels of these innate immunity modulating impurities (IIMIs) that may be present in drug products. However, while it is universally agreed that removal of IIMIs from drug products is crucial for patient safety and to prevent long-term immunogenicity, there is no single assay capable of directly detecting all potential IIMIs or indirectly quantifying downstream biomarkers. Additionally, there is a lack of agreement as to which of the many analytical assays currently employed should be standardized for general IIMI screening. Herein, we review the available literature to highlight cellular and molecular mechanisms underlying IIMI-mediated inflammation and its relevance to the safety and efficacy of pharmaceutical products. We further discuss methodologies used for direct and indirect IIMI identification and quantification.

show abstract

“…Importantly, in this study, downstream processing of five mAbs led to β-glucan levels F I G U R E 4 β-Glucan process-mapping overview. The β-glucan contents (pg/mg of mAb) are shown for five process steps (depth filtration; CEX; VF; UF/DF; drug substance) for five mAbs (1)(2)(3)(4)(5). Detectable β-glucan results are shown as full circles (•).…”

Section: Process-mapping Overviewmentioning

confidence: 99%

Introduction and clearance of beta‐glucan in the downstream processing of monoclonal antibodies

Kluters

Steinhauser

Pfänder

et al. 2021

Biotechnol Progress

View full text Add to dashboard Cite

β-Glucan process-related impurities can be introduced into biopharmaceutical products via upstream or downstream processing or via excipients. This study obtained a comprehensive process-mapping dataset for five monoclonal antibodies to assess β-glucan introduction and clearance during development and production runs at various scales. Overall, 198 data points were available for analysis. The greatest β-glucan concentrations were found in the depth-filtration filtrate (37-2,745 pg/ml). Load volume correlated with β-glucan concentration in the filtrate, whereas flush volume was of secondary importance. Cation-exchange chromatography significantly cleared β-glucans. Furthermore, β-glucan leaching from the Planova 20N virus removal filter was reduced by increasing the flush volume (1 vs. 10 L/m 2 ). β-glucan concentrations after filter flush with 10 L/m 2 were consistently <10 pg/ml. No or only limited β-glucan clearance was attained via ultrafiltration/diafiltration (UF/DF). However, during the first run with monoclonal antibody (mAb) 4, β-glucan concentration in the UF/DF retentate was 10.8 pg/mg, potentially due to β-glucan leaching from the first run with a regenerated cellulose membrane. Overall, β-glucan levels in the final mAb drug substance were 1-12 pg/mg. Assuming high doses of 1,000-5,000 mg, a β-glucan contamination at 20 pg/mg would translate to 20-100 ng/dose, which is below the previously suggested threshold for product safety (≤500 ng/dose). K E Y W O R D S depth filtration, downstream processing, monoclonal antibody, process-related impurities, β-glucan 1 | INTRODUCTION β-Glucans are large polysaccharides with varied chemical structures in which the β-D-glucose monomers are frequently linked by) glycosidic bonds. They occur naturally in the cell walls of bacteria, yeast, cereals, seaweed, and fungi, and have widely varying molecular weights that can range from thousands to millions of Daltons. 1-3 β-(1,3)-D-Glucans are becoming increasingly recognized as pharmaceutical contaminants with immunomodulating properties that have the potential to cause infusion reactions; 1,4 they are commonly described as innate immunity-modulating impurities or process-related impurities (PRI). 1,[4][5][6] During pharmaceutical production of therapeutic proteins such as monoclonal antibodies (mAbs) by mammalian cell culture processes, there are many sources for β-glucan contamination. 1 These sources include cellulose-based filters or membranes, and fungal

show abstract

Detection of Beta-Glucan Contamination in Nanotechnology-Based Formulations

Cited by 17 publications

References 64 publications

Systematic Modification and Evaluation of Enzyme-Loaded Chitosan Nanoparticles

Systematic Modification and Evaluation of Enzyme-Loaded Chitosan Nanoparticles

Innate Immunity Modulating Impurities and the Immunotoxicity of Nanobiotechnology-Based Drug Products

Introduction and clearance of beta‐glucan in the downstream processing of monoclonal antibodies

Contact Info

Product

Resources

About