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Deep-UV Resonance Raman Spectroscopy Enables Direct Quantification of mRNA in Intact Lipid Nanoparticle Formulations.

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Deep-UV resonance Raman spectroscopy (DUV-RRS) rapidly quantifies mRNA in lipid nanoparticles without sample destruction. This label-free method provides accurate concentration measurements for quality control.

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Area of Science:

  • Analytical Chemistry
  • Biotechnology
  • Spectroscopy

Background:

  • Accurate quantification of mRNA within lipid nanoparticles (LNPs) is crucial for therapeutic development.
  • Current methods often require sample destruction (deformulation) and are time-consuming.
  • Developing rapid, label-free analytical techniques is essential for efficient mRNA LNP manufacturing and quality control.

Purpose of the Study:

  • To establish Deep-UV resonance Raman spectroscopy (DUV-RRS) as a rapid, label-free method for quantifying mRNA within intact LNPs.
  • To demonstrate the ability of DUV-RRS to measure mRNA concentrations across various buffers and formulations without deformulation.
  • To assess the potential of DUV-RRS as a high-throughput readout for mRNA LNP quality control.

Main Methods:

  • Utilized Deep-UV resonance Raman spectroscopy (DUV-RRS) for direct, in situ analysis of mRNA within intact LNPs.
  • Employed seconds-scale spectral acquisitions to isolate adenine and guanine Raman signatures.
  • Implemented a multipixel signal-to-noise ratio (SNR) strategy to enhance detection sensitivity, particularly at low mRNA concentrations.
  • Developed calibration models for accurate concentration determination across different formulations and buffer conditions.

Main Results:

  • DUV-RRS successfully quantified mRNA within intact LNPs in seconds, bypassing the need for deformulation.
  • Specific adenine and guanine spectral signatures were isolated with minimal interference from LNP lipid components and water.
  • Calibrated concentration estimates showed accuracy across diverse buffer compositions and LNP formulations.
  • The multipixel SNR strategy significantly improved the detection limits for low mRNA concentrations.

Conclusions:

  • DUV-RRS provides a rapid, label-free, and in situ method for quantifying mRNA in intact LNPs.
  • This technique offers a valuable tool for high-throughput analysis in mRNA formulation development and manufacturing.
  • DUV-RRS streamlines quality control processes for mRNA-based therapeutics, enhancing efficiency and accuracy.