Improved Stability of a Model IgG3 by DoE-Based Evaluation of Buffer Formulations

Formulating appropriate storage conditions for biopharmaceutical proteins is essential for ensuring their stability and thereby their purity, potency, and safety over their shelf-life. Using a model murine IgG3 produced in a bioreactor system, multiple formulation compositions were systematically explored in a DoE design to optimize the stability of a challenging antibody formulation worst case. The stability of the antibody in each buffer formulation was assessed by UV/VIS absorbance at 280 nm and 410 nm and size exclusion high performance liquid chromatography (SEC) to determine overall solubility, opalescence, and aggregate formation, respectively. Upon preliminary testing, acetate was eliminated as a potential storage buffer due to significant visible precipitate formation. An additional 24full factorial DoE was performed that combined the stabilizing effect of arginine with the buffering capacity of histidine. From this final DoE, an optimized formulation of 200 mM arginine, 50 mM histidine, and 100 mM NaCl at a pH of 6.5 was identified to substantially improve stability under long-term storage conditions and after multiple freeze/thaw cycles. Thus, our data highlights the power of DoE based formulation screening approaches even for challenging monoclonal antibody molecules.

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Expired Probiotics: What is Really in Your Cabinet?

10.21203/rs.3.rs-34777/v1 ◽

2020 ◽

Author(s):

Hannah Wilcox ◽

Charles Carr ◽

Shannon Seney ◽

Gregor Reid ◽

Jeremy Burton

Keyword(s):

Storage Conditions ◽

Rrna Gene ◽

Expiry Date ◽

Product Formulation ◽

Long Term Storage ◽

Original Product ◽

Expiration Time ◽

Rrna Gene Sequencing ◽

The Stability

Abstract Background: The popularity of using probiotics has surged, since they became widely accepted as safe and help improve general health. Inevitably, some of these products are used after expiration when microbial cell viability is below the recommended effective dose. Given that probiotics are live microorganisms administered in adequate amounts, the aim of this study was to measure viability in expired products and assess how packaging and storage conditions impact efficacy, if at all.Results: Thirty-three expired probiotic products were evaluated, of which 26 were stored in conditions recommended by the manufacturer. The viable microbial counts were enumerated and representative isolates identified by 16S and ITS rRNA gene sequencing. While the products had a mean past expiration time of 11.32 (1 to 22) years, 22 still had viable contents, and 5 were within or above the original product cell count claim. Product formulation, and number of species present did not appear to impact the stability of the products. However, overall packaging type, storage conditions and time since expiry were found to affect viability. All products with viable cells had the strain stipulated on the label.Conclusion: Despite some selected probiotic products retaining viability long past their expiry date (indicating long term storage is possible), the total counts were mostly well below that required for efficacious use as recommended by the manufacturer. Consuming expired probiotics may not yield the benefits for which they were designed.

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Changes of salivary biomarkers under different storage conditions: effects of temperature and length of storage

Biochemia Medica ◽

10.11613/bm.2019.010706 ◽

2018 ◽

Vol 29 (1) ◽

pp. 94-111 ◽

Cited By ~ 12

Author(s):

Tomás Barranco ◽

Asta Tvarijonaviciute ◽

Damián Escribano ◽

Fernando Tecles ◽

José J Cerón ◽

...

Keyword(s):

Antioxidant Capacity ◽

Room Temperature ◽

Storage Conditions ◽

Trolox Equivalent Antioxidant Capacity ◽

Long Term Storage ◽

Reducing Ability ◽

Trolox Equivalent ◽

The Stability ◽

Term Storage

Introduction: In this report, we aimed to examine the stability of various analytes in saliva under different storage conditions. Materials and methods: Alpha-amylase (AMY), cholinesterase (CHE), lipase (Lip), total esterase (TEA), creatine kinase (CK), aspartate aminotransferase (AST), lactate dehydrogenase (LD), lactate (Lact), adenosine deaminase (ADA), Trolox equivalent antioxidant capacity (TEAC), ferric reducing ability (FRAS), cupric reducing antioxidant capacity (CUPRAC), uric acid (UA), catalase (CAT), advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2) were colorimetrically measured in saliva obtained by passive drool from 12 healthy voluntary donors at baseline and after 3, 6, 24, 72 hours, 7 and 14 days at room temperature (RT) and 4 ºC, and after 14 days, 1, 3 and 6 months at – 20 ºC and – 80 ºC. Results: At RT, changes appeared at 6 hours for TEA and H2O2; 24 hours for Lip, CK, ADA and CUPRAC; and 72 hours for LD, Lact, FRAS, UA and AOPP. At 4 ºC changes were observed after 6 hours for TEA and H2O2; 24 hours for Lip and CUPRAC; 72 hours for CK; and 7 days for LD, FRAS and UA. At – 20 ºC changes appeared after 14 days for AST, Lip, CK and LD; and 3 months for TEA and H2O2. At – 80 ºC observed changes were after 3 months for TEA and H2O2. Conclusions: In short-term storage, the analytes were more stable at 4 ºC than at room temperature, whereas in long-term storage they were more stable at - 80 ºC than at – 20 ºC.

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Long-term stability of ready-to-use 1-mg/mL midazolam solution

American Journal of Health-System Pharmacy ◽

10.1093/ajhp/zxaa040 ◽

2020 ◽

Vol 77 (9) ◽

pp. 681-689

Author(s):

Sixtine Gilliot ◽

Morgane Masse ◽

Frédéric Feutry ◽

Christine Barthélémy ◽

Bertrand Décaudin ◽

...

Keyword(s):

High Performance ◽

Detection Method ◽

Degradation Products ◽

Storage Conditions ◽

Target Concentration ◽

Long Term Stability ◽

Ph Values ◽

Particle Contamination ◽

The Stability

Abstract Purpose Midazolam is a benzodiazepine derivative commonly used in intensive care units to control sedation. Its use requires dilution of a 5-mg/mL commercial solution to a target concentration of 1 mg/mL. A study was conducted to evaluate the stability of diluted ready-to-use 1-mg/mL midazolam solutions over 365 days when stored in cyclic olefin copolymer vials or polypropylene syringes. Methods A specific stability-indicating high-performance liquid chromatography coupled with UV detection method was developed for midazolam hydrochloride and validated for selectivity, linearity, sensitivity, precision, and accuracy. Three storage conditions were tested: –20°C ± 5°C, 5°C ± 3°C, and 25°C ± 2°C at 60% ± 5% relative humidity. Half of the vials were stored upside down to test for the absence of interaction between midazolam and the stopper. Particle contamination, sterility, and pH were assessed. Results The limit of stability was set at 90% of the initial concentration. After 1 year’s storage at –20°C and 5°C, concentrations remained superior to 90% under all storage conditions. At 25°C, stability was maintained up to day 90 in syringes (mean [SD], 92.71% [1.43%]) and to day 180 in upright and upside-down vials (92.12% [0.15%] and 91.57% [0.15%], respectively). No degradation products were apparent, no variations in pH values were detected, and containers retained their sterility and conformity with regard to any specific contamination during the study. Conclusion The evaluated 1-mg/mL midazolam solution was stable over a 1-year period when stored at a refrigerated (5°C) or frozen (–20°C) temperature in both vials and syringes; with storage at 25°C, the stability duration was lower. The preparation of ready-to-use midazolam solutions by a hospital pharmacy is compatible with clinical practice and could help to decrease risks inherent in dilution in care units.

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Untargeted Metabolomics to Evaluate the Stability of Extra-Virgin Olive Oil with Added Lycium barbarum Carotenoids during Storage

Foods ◽

10.3390/foods8060179 ◽

2019 ◽

Vol 8 (6) ◽

pp. 179 ◽

Cited By ~ 12

Author(s):

Domenico Montesano ◽

Gabriele Rocchetti ◽

Lina Cossignani ◽

Biancamaria Senizza ◽

Luna Pollini ◽

...

Keyword(s):

Olive Oil ◽

High Performance ◽

Virgin Olive Oil ◽

Extra Virgin Olive Oil ◽

Untargeted Metabolomics ◽

Lycium Barbarum ◽

Long Term Storage ◽

The Stability ◽

Term Storage

A carotenoid-rich extract from Lycium barbarum L. was added to extra virgin olive oil (EVOO), obtaining a carotenoid-enriched oil (EVOOCar). The oxidative stability of EVOO and EVOOCar was evaluated during long-term storage of 28 weeks at room temperature, by measuring some classical parameters (acidity and peroxide values, spectrophotometric coefficients, fatty acid composition) and the content of minor compounds (i.e., α-tocopherol and lutein). At the end of the storage, higher content (p < 0.01) of α-tocopherol in EVOOCar in respect to EVOO were observed. Zeaxanthin dipalmitate, the most abundant carotenoid compound of Goji berries, decreased slightly (p < 0.05) in EVOOCar until the end of the storage. In regard to polyphenols, an ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC/QTOF-MS) using untargeted metabolomics was carried out. This latter approach discriminated the two oil samples during long-term storage, allowing to identify also the phenolic classes most exposed to significant variations during storage (i.e., mainly lignans and flavones). Besides, the addition of Goji carotenoids preserved the stability of tyrosol equivalents in EVOOCar during long-term storage. These results highlighted that the enrichment of EVOO with a carotenoid-rich extract can improve the shelf-life and nutritional value of added-oil, protecting EVOO natural antioxidants during long-term storage.

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Expired probiotics: what is really in your cabinet?

FEMS Microbes ◽

10.1093/femsmc/xtaa007 ◽

2020 ◽

Author(s):

Hannah Wilcox ◽

Charles Carr ◽

Shannon Seney ◽

Gregor Reid ◽

Jeremy P Burton

Keyword(s):

Storage Conditions ◽

Rrna Gene ◽

Expiry Date ◽

Product Formulation ◽

Long Term Storage ◽

Original Product ◽

Expiration Time ◽

Rrna Gene Sequencing ◽

The Stability

Abstract The popularity of using probiotics has surged, since they became widely accepted as safe and help improve general health. Inevitably, some of these products are used after expiration when microbial cell viability is below the recommended effective dose. Given that probiotics must be live microorganisms administered in adequate amounts, the aim of this study was to measure viability in expired products and assess how packaging and storage conditions impact efficacy, if at all. Thirty-three expired probiotic products were evaluated, of which 26 were stored in conditions recommended by the manufacturer. The viable microbial cells were enumerated and representative isolates identified by 16S and ITS rRNA gene sequencing. While the products had a mean past expiration time of 11.32 (1 to 22) years, 22 still had viable contents, and 5 were within or above the original product cell count claim. Product formulation, and the number of species present did not appear to impact the stability of the products. However, overall packaging type, storage conditions and time since expiry were found to affect viability. All products with viable cells had the strain stipulated on the label. Despite some selected probiotic products retaining viability past their expiry date (indicating long term storage is possible), the total counts were mostly well below that required for efficacious use as recommended by the manufacturer. Consuming expired probiotics may not yield the benefits for which they were designed.

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Human plasma stability during handling and storage: impact on NMR metabolomics

The Analyst ◽

10.1039/c3an02188b ◽

2014 ◽

Vol 139 (5) ◽

pp. 1168-1177 ◽

Cited By ~ 86

Author(s):

Joana Pinto ◽

M. Rosário M. Domingues ◽

Eulália Galhano ◽

Cristina Pita ◽

Maria do Céu Almeida ◽

...

Keyword(s):

Human Plasma ◽

Room Temperature ◽

Storage Conditions ◽

Plasma Composition ◽

Long Term Storage ◽

Short And Long Term ◽

The Stability ◽

And Storage ◽

Term Storage

The stability of human plasma composition was investigated by NMR, considering different collection tubes, time at room temperature (RT), short- and long-term storage conditions and up to 5 consecutive freeze–thaw cycles.

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Investigation of stability of intrauterine aerosol preparation “Yodozol”

Scientific Messenger of LNU of Veterinary Medicine and Biotechnology ◽

10.32718/nvlvet9206 ◽

2018 ◽

Vol 20 (92) ◽

pp. 29-33

Author(s):

R. M. Sachuk ◽

O. A. Katsaraba

Keyword(s):

Shelf Life ◽

Medicinal Product ◽

Storage Temperature ◽

Storage Conditions ◽

Analgesic Effects ◽

Aerosol Preparation ◽

Long Term Storage ◽

The Stability ◽

Term Storage

For standardization, quality control, study of stability and establishment of storage conditions and terms of use, complex preclinical trials of the new development of PE “Biopharm” and the Experimental Station of Epizootology IVM NAAN – aerosol preparation “Yodozol” have been carried out. The methods of evaluation of a medicinal product applied to aerosols are used, which include: determination of changes in appearance, inspection of packaging for leakproofness, measurement of the percentage of contents of the package, the establishment of qualitative and quantitative indicators of active substances, and also the study of microbiological purity of the product. “Yodozol” is a light yellow liquid, 1 ml of which contains 5 mg of iodine and 10 mg of potassium iodide. The drug is used for the prevention and treatment of postnatal intrauterine infections in cows, pigs, sheep and goats (endometritis, pyrometers, cervicitis, vaginitis, delayed digestion caused by microorganisms sensitive to iodine), after obstetrics aid, cesarean section and postpartum sanitation of the uterus. The drug has antimicrobial, anti-inflammatory and analgesic effects, improves the proliferative processes of the genital organs, reduces the time for recovery of animals. The drug is used according to the guidelines, after its production livestock is used without restrictions. The shelf-life, which is the result of the test of the dasg according to the «stability» indicator, has been determined, which was performed under long-term storage in a place protected from light at a temperature range from + 5 ± 2 °С to + 25 ± 2 °С. The studies conducted after 6, 12, 24 and 30 months showed complete compliance of the quality indices with the declared standards when stored for 24 months in the temperature corridor from + 5 °C to + 20 °C. With an increase in storage temperature to + 25 °C or more, a slight quantitative decrease in the concentration of antimicrobial components occurred. In addition, with long-term storage of drugs, release of the contents from the cylinder became uneven and foam acquired a shade less than the saturation rate, increased microbiological contamination. Thus, according to the results of the study, the established shelf life of the preparation is 2 years at the recommended storage temperature from +5 to +20 °С. All studies conducted on the stability of the aerosol intrauterine drug “Yodozol” were included in the registration materials of the medicinal product.

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Isolation and Thermal Stabilization of Bacteriocin Nisin Derived from Whey for Antimicrobial Modifications of Polymers

International Journal of Polymer Science ◽

10.1155/2017/3072582 ◽

2017 ◽

Vol 2017 ◽

pp. 1-7 ◽

Cited By ~ 3

Author(s):

Pavlina Holcapkova ◽

Zuzana Kolarova Raskova ◽

Martina Hrabalikova ◽

Alexandra Salakova ◽

Jan Drbohlav ◽

...

Keyword(s):

Antibacterial Activity ◽

Polyethylene Glycol ◽

High Performance ◽

Thermal Stabilization ◽

Nisin Production ◽

Long Term Stability ◽

Long Term Storage ◽

The Stability ◽

Term Storage

This work describes novel alternative for extraction of bacteriocin nisin from a whey fermentation media and its stabilization by using polyethylene glycol as matrix with high practical applicability. This product was compared with commercially available nisin product stabilized by sodium chloride and nisin extracted and stabilized by using ammonium sulfate and polysorbate 80. The stability of samples was tested by means of long-term storage at −18, 4, 25, and 55°C up to 165 days. The nisin content in the samples was determined by high-performance liquid chromatography and electrophoresis. In addition, effect of whey fortification with lactose on nisin production and antibacterial activity studied against Staphylococcus aureus was tested. Results show that stabilization by polyethylene glycol provides enhanced nisin activity at 55°C after 14 days and long-term stability at 25°C with keeping antibacterial activity.

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The Stability and Efficacy of Tricaine Methanesulfonate (MS222) Solution After Long-Term Storage

Journal of the American Association for Laboratory Animal Science ◽

10.30802/aalas-jaalas-19-000067 ◽

2020 ◽

Author(s):

Erin M Katz ◽

David K Chu ◽

Kerriann M Casey ◽

Katechan Jampachaisri ◽

Stephen A Felt ◽

...

Keyword(s):

Storage Conditions ◽

Specific Storage ◽

Long Term Storage ◽

All Solutions ◽

Clinically Significant ◽

The Stability ◽

Stock Solutions ◽

Term Storage ◽

Tricaine Methanesulfonate

Tricaine methanesulfonate (MS222) is widely used for the anesthesia and euthanasia of laboratory zebrafish. Fresh solutions have been recommended for each use; however, researchers often mix and store concentrated stock solutions for convenience and to reduce occupational exposure and environmental waste. While this is common practice, published guidelines are often inconsistent. Thus, the objective of this study was to evaluate the stability and anesthetic efficacy of MS222 after long-term storage and to develop specific storage parameters. Stock solutions (100 mg/mL MS222) were mixed and stored in amber jars at 4 °C and -20 °C for 2- and 6-mo. Stability of the solutions was analyzed using liquid chromatography-ion trapmass spectrometry and compared with fresh MS222. Fifty adult (30 male, 20 female) wildtype AB zebrafish (Danio rerio) wererandomly anesthetized with 150 mg/L of one of the following MS222 solutions to evaluate anesthetic efficacy: 1) freshly prepared(0m); 2) 2 mo at 4 °C (2m4); 3) 2 mo at -20 °C (2m-20); 4) 6 mo at 4 °C (6m4); 5) 6 mo at -20 °C (6m-20). Time to cessation of swimming, loss of equilibrium, lack of response to von Frey (VF) stimulation, return of equilibrium, and resumption of swimming were compared between groups. Two fish from each group were euthanized at 24-h and 2-wk after anesthesia, and histopathology was performed. All solutions were determined to be stable under all storage conditions. No clinically significant differences were observed between the fresh and stored stock groups during anesthetic testing. No evidence ofanesthetic-related histologic changes were noted in the gills, skin, kidneys, muscle, and central nervous system. Hepatic megalocytosis and a reduction in hepatic vacuolation were seen to varying degrees across all groups, but did not follow a treatment-related trend. Therefore, 100 mg/mL solutions of MS222 can be stored in amber jars at 4 °C or -20 °C for 6 mo and still used to effectively anesthetize zebrafish.

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