mRNA Vaccine Bioanalysis Platform

Rochi Pharmaceutical can provide comprehensive support and services for the safety and effectiveness evaluation of various new drugs and vaccines， specifically the research of LNP-mRNA drugs and vaccines, accumulating rich experience. Hence we establish the bioanalysis platform for mRNA vaccines.
mRNA characteristics:
mRNA vaccine transport mRNA obtained from target gene expression into cells through a specific delivery system. Then, the mRNA will directly express the target protein in the body to stimulate the specific immune response, so that the body can obtain protection from the immune system. mRNA has the below characteristics:
1.Could be introduced into cells to express the corresponding antigen protein in vivo, avoiding the process of protein expression and purification in vitro;
2.Could stimulate the immune system to produce humoral and/or cellular immune responses and play corresponding immune prevention and/or immunotherapy effects;
3.Could 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;
4.Since the degradation of mRNA is accomplished through normal cell metabolism, the potential risk of genetic mutations induced by infection or integration is reduced.
Based on the mechanism and characteristics of mRNA vaccines, Rochi’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：
Rochi equips with 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 can be 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.
Rochi’s Bioanalysis Department has equipped with several M4 Series Microplate Readers and iD5 Series Microplate Readers of Molecular Devices.  The MSD series electrochemiluminescence analyzer supports various types of immunoassay methods and could perform detection and monitoring of different sensitivity ranges of protein antigen levels translated by mRNA.
1.Enzyme-linked Immunoassay (EIA)
2.Fluorescence Immunoassay (FIA)
3.Time-Resolved Fluorescence Immunoassay (TRFIA)
4.Fluorescence Resonance Energy Transfer (FRET)
5.Chemiluminescence Immunoassay (CLIA)
6.Electrochemiluminescence (ECL)
Evaluation of T Cell Factor Secretion Level
In addition to the production of antibodies by the humoral immune response of B cells, the mRNA vaccine can also stimulate the immune response of cells.  The Bioanalysis Department of Rochi has multiple cytokine analysis platforms such as MSD, luminex and FACS CBA to provide sample-saving and cost-saving cytokine analysis technical support.  Rochi even provides the analysis of ELISPOT single-cell level cytokine abundance change.

1.MSD Multi Plex
2.Luminex Multiplex
3.FACS CBA
Analysis of Antibody and Key Domain Antibody Titer
The routine evaluation of antibody titer secreted by B cells is mainly based on the indirect ELISA titer detection method.  The Rochi team devoted to the research of nucleic acid and protein vaccine products in the early years, obtained related patents and received a series of trainings 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 the antibody and key domain antibody titers of the vaccine and evaluate their relevance.  Base on the MSD platform, Rochi’s Bioanalysis Department could also use the validated kit to perform IgG, IgM, IGA inspection of five different SARS-CoV-2 mRNA expression antigens, SARS-CoV-1 Spike, SARS-CoV-2 N, SARS -CoV-2 S1 NTD, SARS-CoV-2 S1 RBD, and SARS-CoV-2.
1.V-PLEX SARS-CoV-2 Panel 1 (IgG)
2.V-PLEX SARS-CoV-2 Panel 2 (IgM)
3.V-PLEX SARS-CoV-2 Panel 3 (IgA)
Neutralizing Antibody Analysis based on Competitive Assay or Pseudovirus
The Rochi vaccine analysis platform could use the electrochemiluminescence MSD platform to apply competitive ligand binding analysis, such as the analysis of functional antibodies stimulated by the expression of antigens in the new 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.