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siRNA-LNP Synthesis

siRNA-LNP Synthesis involves the formulation of custom LNPs for the delivery of small interfering RNA (siRNA) molecules. We employ specialized equipment such as: Lipid Nanoparticle Synthesis Pack, from Elveflow, to produce LNPs with high encapsulation efficiency and low toxicity. The facility also employ the cutting-edge instruments for the siRNA-LNP analysis and characteristic. They include but not limited to: analytical UPLC with PDA, ELS, and MS detectors system are ideal for LNP formulation’s quality and quantitative analysis;   Dynamic Light Scattering (DLS) system are powerful instruments for particles size and zeta potential measurement; quantitative PCR (qPCR) works for a variety of genomic research applications.    

Lipid Nanoparticle Synthesis Pack

(LVF-PAK-LNP-HC-8bar, coming soon!)

The lipid nanoparticle Pack offered by Elveflow is designed for generation of lipid nanoparticles (LNP) using microfluidics techniques. Thanks to the pressure-driven flow control system, a wide range of flow rates (TFR) – from µL/min to 10s of mL/min – can accommodate both low (µL) and large (L) volume production. This versatile platform can be easily scaled up to meet your requirements from the screening to the production stage.

Specifications and Features:

  • Reduced mixing time

  • Increased homogeneity

  • High monodispersity (polydispersity index (PDI) lower than 0.2;

  • Continuous production and high throughput

  • Nanoparticle production integration and automatisation

  • Wide range of monodisperse nanoparticle sizes possible due to precise flow rate control

  • Excellent repeatability from one run to another

  • Possibility of working with both small (μL) and large (L) volumes using the same system.

ACQUITY® UPLC System with PDA eλ, ELS Detectors, or MS Dectector

Ultra performance liquid chromatography (UPLC) is a significant advancement over the conventional high performance liquid chromatography (HPLC) that achieves dramatic increases in resolution, speed and sensitivity in liquid chromatography. The UPLC technology allows users to work at higher efficiencies with a much wider range of linear velocities, flow rates, and backpressures.

Specifications and Features:

  • The PDA eλ detector:

    • Wavelength range: 190 to 800 nm

    • PDA signal linearity range: up to 2.0 AU

    • Simultaneous 2D and 3D operation

    • Able to quantitate trace impurities

  • The ELS detector:

    • 60° scatter angle detection to minimize stray light and optics polarization for long-term optimal sensitivity

    • Nanogram-level detection limits

    • Improved detection for semi-volatile compounds

    • Able to detect lipids

  • Optimized MS/MS detection
    MS ionization options: 

    • Electrospray and multi-mode ionization

    • Atmospheric chemical ionization

    • Dual-mode atmospheric pressure photo-ionization / atmospheric chemical ionization interface;

The facility is equipped with two UPLC systems from Waters. The ACQUITY® UPLC system with PDA eλ and TQD detectors allows users to characterize their formulations with 2D/3D spectroscopy light absorbance as well as their molecular weight. The PDA eλ detector allows users to detect and quantify lower concentrations of sample analytes and compare spectra across a broad wavelength range, while the TQD detector features the highest levels of tandem quadrupole MS selectivity, robustness, speed, and accuracy. The second ACQUITY® UPLC system with PDA eλ, and evaporative light scattering (ELS) and detectors allows users to characterize their formulations with 2D/3D spectroscopy light absorbance, improved lipid detection. The ELS detector complements the PDA eλ detector by providing enhanced sensitivity for the detection of semi-volatile components and low peak dispersion such as lipid-based nanomedicine.

Zetasizer Pro-Red

The Zetasizer Pro-Red is a popular dynamic light scattering (DLS) system from Malvern. It is an easy-to-use system and is commonly used to measure the size (hydrodynamic diameter), polydispersity, molecular weight and zeta potential of the sample.

Measurement modes:

  • Size measurement

  • Zeta potential measurement

  • Molecular weight measurement

  • Kinetics and trend measurements

Specifications and Features:

  • Size range: 0.3nm to 10 um

  • Zeta potential range: -150 to 150 mV

  • Molecular weight range: 1000 to 2 x 107 Daltons

  • Laser wavelength: He-Ne 633 nm

  • Sample volume: 12 to 500 μL for size, 1mL for zeta potential

  • Temperature range: 2 to 90 °C

DelsaMax™ Pro Particle Characterization Instrument

One of the fastest nanoparticle analyzers, the DelsaMax™ Pro from Beckman Coulter allows for the simultaneous analysis of both particle size and zeta potential for sample volumes as small as 45 μl, in less than one second. With an unprecedented 32 detectors working simultaneously, the instrument provides much reduced run time and increased measurement accuracy.

Specifications and Features:

  • Size range: 0.4 to 10,000 nm, hydrodynamic diameter

  • Molar mass range: 5×107 g/mol (Da)

  • Minimum sample volume: 45 μL (170 μL for zeta potential)

  • Minimum measurement time: 1 second

  • Ion strength range: 0 to 50 mS/cm

  • Mobility range: no practical limit

Evaluation of gene expression or gene knock down level Real-Time PCR (qPCR)

Real-time polymerase chain reaction (real-time PCR), or quantitative PCR (qPCR), is a versatile technique that is used to amplify and simultaneously detect and quantify target nucleic acid sequences. 

The StepOnePlus™ real-time PCR system from Applied Biosystems is a 96-well real-time PCR instrument perfect for both first-time and experienced users. With intuitive data analysis and instrument control software, this system is designed to deliver precise, quantitative real-time PCR results for a variety of genomic research applications.

For messenger RNA detection, reverse transcription is required prior to qPCR. The Veriti 96-Well Thermal Cycler from Applied Biosystems is a versatile instrument that provides such feature. In addition, this thermal cycler allows for fast RT-PCR for users who are unable to perform qPCR on their samples.​

Applications:

  • Gene expression analysis

  • Protein expression

  • MicroRNA expression

  • SNP genotyping

  • Viral load analysis

  • Translocation analysis

Specifications and Features for StepOnePlus™ real-time PCR system:

  • For real-time PCR (qPCR) purposes

  • Format: 1 x 96-well plate, 96 x 0.1 mL tubes, 12 x 8-tube strips

  • Reaction volume: 10-30 µL

  • Temperature range: 4-100 °C

  • Run time: < 2 hrs/run in standard mode, < 40 min/run in fast mode

  • Calibrated dye: VIC®, TAMRA™, NED, ROX™, SYBR® Green I, FAM™, JOE™

  • Optics: 4 emission filters, Photodiode, Blue LED excitation source

Specifications and Features for Veriti® 96-Well Thermal Cycler:

  • For RT-PCR or cDNA synthesis purposes

  • Format: 1 x 96-well plate, 96 x 0.2 mL tubes, 12 x 8-tube strips

  • Reaction volume: 10-80 µL

  • Temperature range: 4-99.9 °C

  • Touch screen

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