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Endosafe pts

Manufactured by Charles River Laboratories
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The Endosafe PTS is a rapid test system for the detection and quantification of bacterial endotoxin. It provides accurate and reliable results in a short timeframe, enabling efficient quality control processes.

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Lab products found in correlation

Endosafe, by Charles River Laboratories (2 mentions) Nanodrop 2000, by Thermo Fisher Scientific (2 mentions) Mycoalert plus mycoplasma detection kit, by Lonza (1 mentions) Endosafe pts system, by Charles River Laboratories (1 mentions) Superose 6 16 70 column, by GE Healthcare (1 mentions) Expi293f cells, by Thermo Fisher Scientific (1 mentions) Wyatt dynapro plate reader 2, by Wyatt Technology (1 mentions) Hek293, by American Type Culture Collection (1 mentions) Hiload 26 600 superdex 75, by Cytiva (1 mentions) Protein a affinity chromatography, by Cytiva (1 mentions) Poly 1 c lmw, by InvivoGen (1 mentions) Phosphate buffered saline (pbs), by Charles River Laboratories (1 mentions) Amersham phastsystem, by GE Healthcare (1 mentions) Jem 2100f, by JEOL (1 mentions) Fp 8300, by Jasco (1 mentions) Hpaec pad, by Thermo Fisher Scientific (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Quant it dsdna assay, by Thermo Fisher Scientific (1 mentions) Bira500, by Avidity (1 mentions) Ampicillin, by Roche (1 mentions)

37 protocols using endosafe pts

1

Quantifying Endotoxin in SDVG Samples

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Two SDVGs (6 cm in length) were cut into 4mm2 pieces and incubated in separate 15 mL conical tubes with 1 mL of 1× PBS, pH 7.4, at 37 °C for 24 h. An Endosafe® endotoxin testing system (Endosafe®-PTS, Charles River, USA) was conducted using the 1× PBS supernatant after centrifugation (500 × g for 5 min) and dilution (1:20) in sterile and endotoxin-free dH2O. The concentration of endotoxin in the eluent (1× PBS) was quantified using the Endosafe® LAL Cartridges (#PTS2005F, Charles River, USA) and the portable system Endosafe®-PTS (Charles River, USA) and expressed in EU mL−1.
Akentjew T.L., Terraza C., Suazo C., Maksimcuka J., Wilkens C.A., Vargas F., Zavala G., Ocaña M., Enrione J., García-Herrera C.M., Valenzuela L.M., Blaker J.J., Khoury M, & Acevedo J.P. (2019). Rapid fabrication of reinforced and cell-laden vascular grafts structurally inspired by human coronary arteries. Nature Communications, 10, 3098.
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2

Detecting Mycoplasma and Endotoxins

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The detection of non-cellular impurities was carried out in accordance with the methodology recommendations of Chapter 2.6.21 and 2.6.7 of the European Pharmacopeia (Eu Ph) for mycoplasma and Chapter 2.6.14 for endotoxins. A DNA-binding dye-based qPCR system was employed for the detection of mycoplasma DNA in cell cultures. The assay was developed by the Genomics Unit in collaboration with the Monoclonal Antibodies Unit, both from CNIO, to detect 16s rRNA gene sequences from up to 70 Mollicutes species. Both specificity and sensitivity were extensively tested through benchmarking with established commercial systems (MTC-NI System, Millipore/GEN-PROBE Cat. No. 4573 and MycoAlert™ PLUS Mycoplasma Detection Kit, Lonza Cat. No. LT07-705). The Clinical Microbiology and Parasitology Service of HULP carried out the endotoxin test Endosafe-PTS (Charles River) to quantify endotoxin levels at day +8 and in final products.
Fernández L., Fernández A., Mirones I., Escudero A., Cardoso L., Vela M., Lanzarot D., de Paz R., Leivas A., Gallardo M., Marcos A., Romero A.B., Martínez-López J, & Pérez-Martínez A. (2019). GMP-Compliant Manufacturing of NKG2D CAR Memory T Cells Using CliniMACS Prodigy. Frontiers in Immunology, 10, 2361.
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3

Polysialic Acid Characterization Protocol

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During cultivation, downstream processing and for the final characterization of the product several analysis were performed. The polySia concentration was measured with a modified thiobarbituric acid assay, protein concentration was determined by the Bradford method, DNA concentration by UV–vis absorption (NanoDrop 2000, Thermo Scientific, Waltham, USA), endotoxin concentration with Endosafe-PTS™ system (Endosafe-PTS™, Charles River Laboratories, Boston, USA) and the chain length characterization with DMB-HPLC analysis (DMB = 1,2-diamino-4,5-methylenedioxybenzene). Cell disruption was performed with ultrasonic treatment. All methods were performed as previously reported [11 ].
de Vries I., Schreiber S., Boßmann D., Hellmann Z., Kopatz J., Neumann H, & Beutel S. (2018). Single-use membrane adsorbers for endotoxin removal and purification of endogenous polysialic acid from Escherichia coli K1. Biotechnology Reports, 17, 110-116.
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4

Endotoxin Quantification Procedure

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Endotoxin was determined with the Endosafe-PTS (Charles-River) according to manufacturer’s protocol. Purified protein preparations containing <1 unit/ml endotoxin were used for experiments.
Kusters D.H., Chatrou M.L., Willems B.A., De Saint-Hubert M., Bauwens M., van der Vorst E., Bena S., Biessen E.A., Perretti M., Schurgers L.J, & Reutelingsperger C.P. (2015). Pharmacological Treatment with Annexin A1 Reduces Atherosclerotic Plaque Burden in LDLR-/- Mice on Western Type Diet. PLoS ONE, 10(6), e0130484.
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5

Recombinant Tc24 and TSA-1 Protein Expression

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The recombinant protein Tc24 (24.7 kDa) was expressed and purified as previously described [23 (link)]. Briefly, the Tc24 coding sequence subcloned into the yeast expression vector pPICZα was used for expression in Pichia pastoris X-33 and the recombinant protein was purified by affinity chromatography. For TSA-1, the coding sequence was subcloned into E. coli expression vector pET41a and purification was achieved by Ni-affinity chromatography under denaturing condition and then refolded by size exclusion chromatography. Endotoxin levels were measured with the Charles River Endosafe-PTS. The integrity and size of the recombinant proteins were analyzed by SDS-PAGE electrophoresis following purification and just before use, after storage at -80° C.
Villanueva-Lizama L.E., Cruz-Chan J.V., Aguilar-Cetina A.D., Herrera-Sanchez L.F., Rodriguez-Perez J.M., Rosado-Vallado M.E., Ramirez-Sierra M.J., Ortega-Lopez J., Jones K., Hotez P., Bottazzi M.E, & Dumonteil E. (2018). Trypanosoma cruzi vaccine candidate antigens Tc24 and TSA-1 recall memory immune response associated with HLA-A and -B supertypes in Chagasic chronic patients from Mexico. PLoS Neglected Tropical Diseases, 12(1), e0006240.
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6

Purification and Characterization of Recombinant HA-NP Nanoparticles

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The SIB-0002 mammalian expression plasmid [13 (link)] containing the A/New Caledonia/20/1999 (NC99) HA-NP gene was transfected into Expi293F cells per manufacturer’s protocol (Thermo), and supernatant was harvested five to six days later. HA-NPs were purified by anion exchange chromatography with an HP-Q column (GE Healthcare) followed by size exclusion chromatography (SEC, Superose 6 16/70 column, GE Healthcare). Correct assembly and size were confirmed by SEC retention volume, SDS-PAGE, and dynamic light scattering (DLS, Wyatt DynaPro Plate Reader II). Endotoxin levels were measured by Endosafe PTS with LAL cartridges (Charles River Laboratories).
Rudicell R.S., Garinot M., Kanekiyo M., Kamp H.D., Swanson K., Chou T.H., Dai S., Bedel O., Simard D., Gillespie R.A., Yang K., Reardon M., Avila L.Z., Besev M., Dhal P.K., Dharanipragada R., Zheng L., Duan X., Dinapoli J., Vogel T.U., Kleanthous H., Mascola J.R., Graham B.S., Haensler J., Wei C.J, & Nabel G.J. (2019). Comparison of Adjuvants to Optimize Influenza Neutralizing Antibody Responses. Vaccine, 37(42), 6208-6220.
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7

Quantifying Endotoxin Levels by PTS

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The vehicle control, VYC-15L fragments and HYC-24L + fragments were quantified by Endosafe PTS (Charles River). The Endosafe®-PTS (Portable Test System) is a rapid, point-of-use test system, which is comprised of a test cartridge along with a hand-held spectrophotometer. For this test, we used cartridge with 0.01 EU/ml sensitivity (PTS2001F). All experiments were carried out according to the manufacturer's instructions.
Sanchez B., Ferraro S., Josset-Lamaugarny A., Pagnon A., Hee C.K., Nakab L., Sigaudo-Roussel D, & Fromy B. (2023). Skin Cell and Tissue Responses to Cross-Linked Hyaluronic Acid in Low-Grade Inflammatory Conditions. International Journal of Inflammation, 2023, 3001080.
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8

Limulus Amebocyte Lysate Endotoxin Test

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After cell isolation, separation from the antibody-conjugated beads and centrifugation, supernatant was diluted 1:10 with Endosafe (Charles River, Charleston, SC, USA) limulus amebocyte lysate reagent water and transferred into an Endosafe PTS cartridge (0.05 EU/mL), which was loaded into an Endosafe PTS reader.
Kerstan A., Niebergall-Roth E., Esterlechner J., Schröder H.M., Gasser M., Waaga-Gasser A.M., Goebeler M., Rak K., Schrüfer P., Endres S., Hagenbusch P., Kraft K., Dieter K., Ballikaya S., Stemler N., Sadeghi S., Tappenbeck N., Murphy G.F., Orgill D.P., Frank N.Y., Ganss C., Scharffetter-Kochanek K., Frank M.H, & Kluth M.A. (2020). Ex vivo-expanded highly pure ABCB5+ mesenchymal stromal cells as Good Manufacturing Practice-compliant autologous advanced therapy medicinal product for clinical use: process validation and first in-human data. Cytotherapy, 23(2), 165-175.
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9

Endotoxin Assessment of Hyaluronic Acid

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HA powder was opportunely dissolved in pyrogen free water. Limulus Amebocyte Lysate (LAL) test was performed to measure the amount of pyrogens (bacterial endotoxins) in the solution. ENDOSAFE®-PTS cartridge US License N.1197 by Charles River Endosafe was used. All operations were performed under conditions avoiding endotoxin contamination. Results were reported as endotoxin units (EU/mg) of HA powder.
Huang L., Wang Y., Liu H, & Huang J. (2017). Local injection of high-molecular hyaluronan promotes wound healing in old rats by increasing angiogenesis. Oncotarget, 9(9), 8241-8252.
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10

Quantifying GRFT Endotoxin Levels

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Contaminating endotoxin levels in the final GRFT product purified at industrial pilot scale was analyzed with Charles River Laboratory Endosafe® PTS. Twenty-five μL of GRFT solution was added to each of the four wells on a Limulus Amebocyte Lysate Test cartridge and the endotoxin levels were analyzed accordingly.
Fuqua J.L., Wanga V, & Palmer K.E. (2015). Improving the large scale purification of the HIV microbicide, griffithsin. BMC Biotechnology, 15(1), 12.
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