mRNA vaccine bioanalysis techniques

Jiangxi Chiyan can provide long-term support for the safety and efficacy evaluation of all kinds of new biotechnology drugs and vaccines, devoting itself to the analysis of the efficacy and safety evaluation of LNP-mRNA drugs and vaccines, we have accumulated abundant experience and established and perfected the related biological analysis technology of the new type of mRNA vaccine.

mRNA characteristics

mRNA vaccine is a kind of vaccine which is prepared by transcription and synthesis of exogenous target gene sequence and introduced into body cells through a specific delivery system to directly express target protein in vivo, stimulate the body to produce specific immunological reaction, to make the body obtain immune protection.

mRNA has the following characteristics: 1.It can be introduced into cells and express the corresponding antigen protein in vivo, avoiding the process of protein expression and purification in vitro. 2.Can stimulate the immune system to produce humoral or cellular immune response, play the corresponding role of immune prevention or immunotherapy; 3.Can stimulate the body's immune system to produce a variety of cytokines and other ways to enhance the body's immune response or change the type of immune response; 4.This type of vaccine has its unique advantages because the degradation of mRNA is accomplished through normal cellular metabolism, reducing the potential risk of gene mutations induced by infection or integration. 

Based on the mechanism and characteristics of mRNA vaccine, Jiangxi Chiyan Technology Pharmaceutical Analysis Department has established a comprehensive framework covering metabolism and biodistribution to support the safety and efficacy evaluation of mRNA vaccine, to the serial evaluation techniques of total antibody, key sequence domain antibody and anti-virus neutralizing antibody titers, as well as cellular immune response efficacy.

Biodistribution analysis of mRNA level

Jiangxi Chiyan cooperates with ABI7500 series Real-Time PCR amplification instrument and supporting Kingfisher nucleic acid extractor equipment, which can perform mRNA metabolism and distribution analysis through subsequent RT-qPCR technology based on Trizol preserved whole blood samples and other types of tissue samples collected by RNAase free tube.

Analysis of biological distribution of mRNA expression protein antigens

mRNA vaccines need to be translated into proteins in the body to stimulate further immune responses, so it is particularly important to confirm mRNA expression levels and distribution at the protein level. The Biological Drug Analysis Department of Jiangxi Chiyan has cooperated with a number of Molecular Device M4 series enzyme marker, new iD5 series enzyme marker and MSD series electrochemical luminescence analyzer, which support various types of immunoassay methods, and can perform detection and monitoring of different sensitivity ranges of mRNA translated protein antigen levels.

1. Enzyme-linked Immunoassay (EIA) 

2. Fluorescence Immunoassay (FIA) 

3. Time Resolved Fluorescence (TRFIA) 

4. Fluorescence Resonance Energy Transfer (FRET)

5. Chemiluminescence (CLIA) 

6. Electrochemical Luminescence (ECL)

Evaluation of T cytokine secretion level

MRNA vaccines not only induce humoral immune responses in B cells to produce antibodies, but also stimulate cellular immune responses. The drug analysis Department of Jiangxi Chiyan Biotechnology has multiple cytokine analysis platforms such as MSD, Luminex, FACS CBA, etc., to provide sample saving and cost saving cytokine analysis technical support. Even ELISPOT single cell level cytokine abundance change analysis.

1. MSD Multi Plex

2. Luminex Multiplex

3. FACS CBA

Titer analysis of total antibody and key domain antibody

The conventional titer evaluation of antibody secreted by B cells is mainly based on indirect ELISA titer detection method. The core members of Jiangxi Zhiresearch Cooperation technical team were committed to nucleic acid and protein vaccine product research in their early years, and received a series of training on vaccine titer biological analysis and evaluation, and established a series of method evaluation system and standard formulation rules based on practical experience. It is used to investigate the total antibody and the titer of the antibody in the key domain of the vaccine and to evaluate their correlation. Based on the MSD platform, Jiangxi Chiyan Biotechnology drug analysis department can also directly perform IgG, IgM, IGA detection of five different important mRNA expression antigens, ie. SARS-CoV-1 Spike, SARS-CoV-2 N, SARS-CoV-2 S1 NTD, SARS-CoV-2 S1 RBD, and SARS-CoV-2 Spike by using validated kits which is based on 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 Assay based on competitive assay or pseudo virus 

The vaccine analysis technology platform can not only apply competitive ligand binding analysis methods based on the electroluminescence MSD platform to perform functional antibody analysis stimulated by the antigen expressed in the COVID-19 mRNA vaccine, i.e. neutralizing antibody analysis, but also have independent cell houses and other facilities to perform neutralizing antibody analysis based on the interaction between fake virus and ACE2 cell lines.