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mRNA characteristics- mRNA vaccine transport mRNA obtained from target gene expression into cells through a specific delivery system. The mRNA can drive the expression of target proteins in the body to stimulate the specific immune response, so that the immune system can be activated. mRNA has the below characteristics:Can be introduced into cells to express the corresponding antigen protein in vivo, avoiding the process of protein expression and purification in vitro;Can stimulate the immune system to produce humoral and/or cellular immune responses driving prevention and/or immunotherapy effects;Can enhance the body’s immune response ability or change the type of immune response by stimulating the body’s immune system to produce a variety of cytokines;Since the degradation of mRNA is accomplished through normal cell metabolism, the potential risk of genetic mutations induced by infection or integration is reduced.Therefore, the mRNA vaccine has unique advantages.
Based on the mechanism and characteristics of mRNA vaccines, Medicilon’s Bioanalysis Department has established an evaluation platform covering metabolism and biodistribution, key sequence domain antibodies and antiviral neutralizing antibody titers and the evaluation of effectiveness of cellular immune response.
mRNA Biodistribution Analysis- Medicilon uses the ABI7500 Real-Time PCR System and the Kingfisher Nucleic Acid Extractor. The blood samples stored in Trizol and other types of tissue samples collected from RNAase free tubes are used for mRNA metabolism and distribution analysis by subsequent RT-qPCR technology.
Biodistribution Analysis of mRNA Expression Protein Antigen- The mRNA vaccine needs to be expressed to stimulate the body for immune response, so it is particularly important to confirm the expression level and distribution of mRNA at the protein level.Medicilon’s Bioanalysis Department is equipped with M4 Series Microplate Readers and iD5 Series Microplate Readers. The MSD series electrochemiluminescence analyzer supports various types of immunoassay methods and can perform detection and monitoring of different sensitivity ranges of protein antigen levels translated by mRNA.Enzyme-linked Immunoassay (EIA)Fluorescence Immunoassay (FIA)Time-Resolved Fluorescence Immunoassay (TRFIA)Fluorescence Resonance Energy Transfer (FRET)Chemiluminescence Immunoassay (CLIA)Electrochemiluminescence (ECL)
Evaluation of T Cell Factor Secretion Level- The Bioanalysis Department of Medicilon has multiple cytokine analysis platforms such as MSD, luminex and FACS CBA to provide sample-saving and cost-saving cytokine analysis technical support. Medicilon also provides the analysis of ELISPOT single-cell level cytokine abundance change.MSD Multi PlexLuminex MultiplexFACS CBA
Analysis of Antibody and Key Domain Antibody Titer- The routine evaluation of antibody titersis undertaken by indirect ELISA titer detection. The Medicilon team has obtained related patents on the biological analysis and evaluation of vaccine potency in Pfizer in the United States. Based on practical experience, a series of method evaluation systems and standard-setting rules have been established to investigate vaccine-derived antibody titers and evaluate their relevance. Medicilon’s Bioanalysis Department can also perform IgG, IgM and IGA evaluations demonstrated on SARS proteins.V-PLEX SARS-CoV-2 Panel 1 (IgG)V-PLEX SARS-CoV-2 Panel 2 (IgM)V-PLEX SARS-CoV-2 Panel 3 (IgA)
Neutralizing Antibody Analysis based on Competitive Assay or Pseudovirus- Medicilon can use the electrochemiluminescence MSD platform to undertake competitive ligand binding analysis, for e.g. analysis of functional antibodies stimulated by the expression of antigens in a coronavirus mRNA vaccine. There are also facilities such as independent cell rooms for neutralizing antibody analysis based on the interaction between pseudoviruses and cell lines ACE2.
