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Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
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Microtiter Plate-Based Differential Scanning Fluorimetry: A High-Throughput Method for Efficient Formulation

Meifeng Nie1, Yue Liu1, Xiaofen Huang1

  • 1State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, Fujian 361102, China.

Journal of Pharmaceutical Sciences
|May 23, 2022
PubMed
Summary
This summary is machine-generated.

A new microtiter plate-based differential scanning fluorimetry (DSF) method accurately assesses protein thermal stability in vaccine adjuvants. This higher throughput technique requires less sample, aiding early-stage vaccine formulation development.

Keywords:
Conformational stabilityDifferential scanning fluorimetryParticulate adjuvantProtein antigenVaccine formulation

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Area of Science:

  • Vaccine Adjuvant Development
  • Protein Biophysics
  • Analytical Chemistry

Background:

  • Nano/microparticles are crucial vaccine adjuvants, enhancing antigen stability and immune responses.
  • Assessing protein conformational stability is vital for vaccine formulation and product quality.
  • Differential Scanning Calorimetry (DSC) is a standard method but has limitations in throughput and sample volume.

Purpose of the Study:

  • To develop and optimize a high-throughput microtiter plate-based differential scanning fluorimetry (DSF) method.
  • To assess protein thermal stability specifically for antigens adsorbed onto particulate adjuvants.
  • To provide a more efficient alternative to DSC for early-stage vaccine formulation.

Main Methods:

  • Development and optimization of a microtiter plate-based differential scanning fluorimetry (DSF) assay.
  • Testing the method using recombinant human papillomavirus (HPV) vaccine antigens and model proteins.
  • Adsorption of proteins onto nano/microparticle adjuvants prior to thermal stability assessment.

Main Results:

  • The plate-based DSF method demonstrated enhanced sensitivity for assessing protein thermal stability.
  • Results showed strong correlation with the established differential scanning calorimetry (DSC) method.
  • The DSF method required significantly less sample (1/10–1/20 of DSC) and offered higher throughput.

Conclusions:

  • Plate-based DSF is a validated, sensitive, and efficient method for evaluating protein thermal stability in particulate adjuvant-adsorbed forms.
  • This method is particularly advantageous for early-stage vaccine formulation development due to reduced sample requirements and increased throughput.
  • The optimized DSF assay provides a valuable tool for enhancing vaccine formulation and quality control processes.