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Lyobeta 15

Manufactured by Telstar
Sourced in Spain

The LyoBeta 15 is a laboratory freeze-dryer designed for efficient lyophilization of samples. It features a temperature-controlled chamber and a high-performance refrigeration system to facilitate the freeze-drying process.

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14 protocols using lyobeta 15

1

Optimized Pressurized Liquid Extraction of Truffles

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According to the results obtained from the response surface plots and PCA analysis in a previous study [11 (link)], the combination of 180 °C and 16.7 MPa applied during 30 min were the optimal PLE conditions for Tuber aestivum and Terfezia claveryi truffles, although extractions from Terfezia truffles were more effective than from the Tuber genus. Thus, the optimized PLE method was applied to seven other truffles species.
Truffle powders (0.5 g) were submitted to pressurized liquid extraction using an Accelerated Solvent Extractor (ASE) (Dionex Corporation, ASE 350, USA) [6 (link)]. Briefly, samples were submitted to 16.7 MPa at 180 °C for 30 min. Fractions obtained with water were immediately frozen and freeze-dried in a LyoBeta 15 lyophilizer (Telstar, Madrid, Spain), and those obtained with ethanol were dried using a rotary vacuum evaporator at 40 °C (IKA® RV 10, VWR International, Barcelona, Spain). The water:ethanol extracts were dried and then lyophilized. Afterwards, samples were stored in darkness at −20 °C.
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2

Truffle Harvesting and Preparation

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Terfezia claveryi truffles were harvested from the experimental station at the University of Murcia (Espinardo, Spain), and Tuber aestivum fruiting bodies were harvested in Gúdar-Javalambre forests (Teruel, Spain). Fresh truffles were identified, selected and processed according to Rivera et al. (2011) [26 (link)]. After, truffle samples (500 g) were lyophilized (LyoBeta 15 lyophilizer (Telstar, Madrid, Spain)), ground, mixed and sieved until a particle size lower than 0.5 mm was obtained. Powdered truffles were kept frozen (at −80 °C) until further use.
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3

Sage Extract Production Protocol

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Two SEs from selected sage plants were manufactured in the laboratory of the Rain Feed Crops for the Rural Development Department part of the Institute for Agrifood and Environmental Research and Development, Murcia, Spain (IMIDA). Essential oil was previously removed from the sage leaves by distillation with water steam, according to the method described by Jordán et al. (2013) [41 (link)]. For the SEs, oil-free dry leaves were ground to 2 mm to obtain a powder. Sage powder was mixed with distilled water at 1:10 (w:v) ratio, kept at 30 °C for 90 min in a water bath with constant stirring and centrifuged at 4560× g and 5 °C for 10 min in a Digecen 21 R centrifuge (Orto Alresa, Madrid, Spain). The supernatant was filtered on Whatman filter paper (No. 4), lyophilised (Lyobeta 15, Telstar) at 100 mbar and −80 °C for 24 h. The dry SEs were packed into a dark steel container with nitrogen and kept at −80 °C and in darkness until further use.
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4

Protein-rGO Binding Analysis by Spectroscopy

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The binding of each protein to rGO surface was studied by Raman and Fourier transform infrared spectroscopy. First, 250 µg/mL rGO was suspended either in 50 µg/mL BSA, 200 µg/mL collagen, or 50 µg/mL elastin solution, and agitated at 37 °C for 2 h. The supernatants were collected after spinning down at rpm for 15 min and lyophilized in a Telstar Lyobeta 15 lyophilizer. rGO without protein incubation and proteins without rGO were also studied. At least three samples were studied for each condition.
Raman spectrum was obtained by confocal Raman imaging (Alpha 300 M, Company WITec, Ulm, Germany) with a 532 nm laser (5% laser power, a contact time of the 50 s, and four accumulations). FT-IR, using an attenuated total reflectance (ATR) technique, was performed in a FT-IR Bruker IFS 66/S Spectrometer, with 32 scans at a resolution of 4 cm−1 between the wavelength ranges of 4000–400 cm−1. Air background was applied as a blank. At least three samples were analyzed for each condition.
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5

Preparation of Chitosan Particles with Marjoram Extract

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Chitosan particles were obtained by inducing the gelation of a chitosan solution with TPP, following the method described by Da Silva et al. [16 (link)], with adaptations. Once the LCH or the MCH (1% w/v) were dissolved in the acetic acid solution (1% v/v), marjoram extract was added at different final concentrations (3, 2, 1, 0.5, and 0.25 mg/mL) and the pH value was adjusted to 5.8 with 1 M sodium hydroxide. Then, TPP was dissolved in milli-Q water at 0.1% (w/v) and, in order to study the best ratio for chitosan:TPP, different volumes of the TPP solution were added to the chitosan solution under magnetic stirring at room temperature for 20 min. The ratios of chitosan:TPP assayed were 5:1, 6:1, and 7:1. After that, the suspension was centrifugated (40,000× g, 45 min, T = 25 °C) and the supernatant was discarded to obtain the particles. The particles were frozen, then freeze-dried (LyoBeta 15, Telstar, Spain) and stored at 4 °C until analyses were performed.
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6

Fourier Transform Infrared Spectroscopy Analysis of GM-SLN

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For the determination of interactions between the components of the GM-SLN vector, Fourier Transform Infrared Spectroscopy (FT-IR) analysis was performed. SLNs and the GM-SLN vector were lyophilized at −50 °C and 0.2 mbar for 42 h (LyoBeta 15, Telstar, Spain) and the powders were mixed homogeneously with potassium bromide. The blends were compressed using a hydraulic compressor applying a pressure of 10 tons for 5 min. The discs obtained were placed in an FT-IR spectrometer (Thermo Fisher Scientific, Madrid, Spain) and the IR spectrums were recorded in the mid-IR region (4000–400 cm−1). The recorded signals were reported as transmittance percentages.
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7

PLGA Nanoparticles for Melanoma Antigen Delivery

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PLGA (Resomer-RG503; lactide:glycolide ratio of 50:50; MW 40,600; viscosity 0.41 dL/g; Evonik, Germany), PEI (branched form, molecular weight of 25,000 Da), and polyvinyl alcohol (PVA) were purchased from Sigma Aldrich, Darmstadt, Germany.
PEI-coated PLGA NPs were prepared using the solvent extraction–evaporation double emulsion (w/o/w) method, with encapsulation of melanoma patients’ antigens. Briefly, 50 mg of PLGA and 0.65 mg of PEI were dissolved in 1 mL of dichloromethane (DCM) and emulsified with a 1.25% (w/v) peptide solution in acetonitrile (ACN):water (50:50) by sonication (30 s using Branson sonifier 250). This emulsion was then mixed with 5 mL of 5% (w/v) PVA and sonicated for 1 min. The final emulsion was poured into a 2% (v/v) isopropanol solution (20 mL) and stirred for 2 h to allow solvent evaporation. Finally, the resulting nanoparticles were freeze-dried for 42 h with trehalose used as a cryoprotectant (Lyobeta 15, Telstar, Barcelona, Spain).
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8

Insect Preparation for Animal Feed

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A. domesticus (adult) and T. molitor (larva), were purchased under frozen conditions in a local company specialized in insect production intended for animal feed (Animal Center SL, Valencia, Spain). Afterwards, insects were gently rinsed with distilled water, freeze-dried (LyoBeta 15, Telstar, Terrasa, Spain), ground (Knife Mill Grindomix GM 200, Retsch GmbH, Haan, Germany) , and stored at -20ºC protected from oxygen, light and moisture until further use.
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9

Peptide-Loaded PLGA Nanoparticles Synthesis

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NPs were prepared by the solvent extraction-evaporation of a double emulsion (w/o/w) method. Briefly, 80 mg of PLGA (50:50 lactide-glicolide ratio. Resomer-RG503; MW 40,600; viscosity 0.41 dl/g. Evonik, Germany) and 1.04 mg of PEI (polyethylenimine, branched form with molecular weight 25,000Da. Sigma Aldrich, Germany) were dissolved in 1.6 ml of DCM (dichloromethane) and emulsified with 80 µL of each peptide suspension in miliQ water using 30 seconds of sonication with a Branson sonifier 250 in an ice bath to avoid high temperatures. The resulting w/o emulsion was then mixed with 8 ml of 5% (w/v) PVA (polyvinyl alcohol. Sigma Aldrich, Germany) and sonicated for an additional 1 minute. Finally, the w/o/w emulsion was poured into a 16 ml 2% (v/v) isopropanol solution and stirred for 2 hours to allow for solvent evaporation. The resultant NPs were washed 3 times in miliQ water and freeze-dryed with 15% (w/w) trehalose for 42 h (Lyobeta 15, Telstar ® ).
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10

Extraction and Characterization of Rosemary Leaf Extract

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Selected high-yielding rosemary plants from a germplasm bank were cultivated and harvested in the experimental farm of the Institute of Agricultural and Food Research and Development (IMIDA, Murcia, Spain). RLE was obtained from the leaf by-product that had been previously distilled with water steam and extracted with water to obtain the essential oil and the RWE, respectively [27 (link)]. Then, 3 g of rosemary by-product were mixed with 450 mL acetone:water (1:1, v/v) and kept under constant stirring (30 °C for 90 min). The mix was centrifuged in a Digecen 21 R centrifuge (Orto Alresa, Madrid, Spain) (4560× g and 5 °C for 10 min) and the supernatant was then filtered (Whatman No. 4). The acetone fraction was removed by applying a vacuum at 40 °C in a Syncore Polyvap R-96 evaporation system (Buchi, Flawil, Switzerland), and the water fraction was removed using a lyophilisation system (Lyobeta 15, Telstar, Terrassa, Spain) (100 mbar and −80 °C for 24 h). Lyophilized RLE, a yellow-greenish dry powder with an intense herbal flavour, was stored at −80 °C until further use.
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