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Yeasen Biotechnology GMP Grade mRNA In Vitro Synthesis Raw Materials

With the rapid development of biotechnology, mRNA vaccines prepared by in vitro RNA synthesis technology have a huge impact on the prevention of COVID-19 infectious diseases. Many companies, such as Moderna in the United States, CureVac in Germany, and BioNTech in Germany, have developed therapeutic mRNA vaccines and drugs for other diseases such as tumors, infectious diseases, and chronic diseases.

The research and preparation of mRNA vaccines require a variety of raw materials to ensure the successful development of vaccines. As an innovative leader in the domestic molecular enzyme industry, Yeasen Bio has successfully developed various molecular enzymes and reagents through the advanced molecular enzyme two-way technology platform and large-scale protein fermentation and purification technology platform. 

To ensure product quality and production capacity, Yeasen Bio has built a GMP-level production base in Wuhan to fully meet customers' industrial production needs, with an annual production capacity of more than 5 billion servings. Also, Yeasen is the first company to receive the ISO 13485 certificate for the research, development, manufacture, and distribution of molecular enzymes in China, which can ensure all reagents quality, consistency, and traceability. 

 

1. Advantage of GMP
2. Yeasen can provide GMP-grade Products
3. Yeasen Acquired DMF Numbers for Many Products
4. Application scenarios of Yeasen products
5. Raw materials for mRNA vaccine preparation provided by Yeasen
6. Ordering Information

 

1. Advantage of GMP

"GMP-grade" is a branding term that Yeasen uses to describe reagents manufactured at ISO 13485 certified facilities. Yeasen’s GMP-grade reagents are produced in compliance with ISO 13485 quality management system standards and with more stringent process controls and complete documentation records. Yeasen can not only offer high-quality reagents but also provide documents, site audits, and other support to our customers.

Table 1. Advantage of GMP

Quality Controls

Research level

GMP-grade

Animal free

Animal-derived material may be used

Animal-free certificates can be provided if required

Cell bank characterization

Less stringent

Stringent (compliant with GMP regulations)

Traceability information for raw materials and finished product

Less stringent

Stringent (compliant with GMP regulations)

Endotoxin level control

/

Stringent

Sterile control

/

Stringent

Batch records for manufacturing and testing

Less stringent

Stringent (Batch records can beh provided if required)

Process validation report

/

The report can be provided if required

Change control system

Self-assessment

Stringent (compliant with GMP regulations)

Quality management system

Self-assessment

ISO 13485 certification

DMF number

/

Yes (reporting)

 

2. Yeasen can provide GMP-grade Products

Based on wide experience and technical advantages in enzyme development and industrial production, Yeasen constructed a new facility, named "mRNAtools", to supply raw materials for mRNA-based drugs. Covering 50000 square feet, the mRNAtools facility is built and operated by GMP regulations and is equipped with 2×1500L automatic fermentation equipment, industrial-scale purification, and lyophilization equipment. To provide more strict reagents and also satisfy the production and registration requirements of customers. The molecular enzyme production base specially built for GMP-grade products ensures the quality of products from both hardware and software. 70 professional production team, team members have many years of GMP production experience. The warehousing and logistics are modularized. While ensuring the safety of product storage and transportation multi-center distribution across the country ensures the supply cycle.

 

3. Yeasen Acquired DMF Numbers for Many Products

Drug master files (DMFs) are documents submitted voluntarily to the US Food & Drug Administration (FDA) that contain confidential, detailed information about facilities, processes, or articles used in the manufacturing, processing, packaging, and storing of human drug products. 
The FDA reviews the technical contents of a DMF when an API manufacturer incorporates it as a reference to support it's Investigational New Drug Application (IND), New Drug Application (NDA), Abbreviated New Drug Application (ANDA), and Export Application. DMFs have significant roles in the application. The FDA approval process is significantly shortened when using raw materials from manufacturers that have filed DMFs for these specific materials. 
Yeasen has got the DMF files that can speed up the project schedule. You may request that we provide reference authorization to our DMFs in support of your application with the FDA. To initiate the reference authorization of our DMFs, please submit a Letter of Authorization request to Yeasen requesting that we provide a DMF Letter of Authorization to the respective FDA Center. 

 

4. Application scenarios of Yeasen products

4.1 Template Generation

Plasmid linearization is an indispensable step during template generation when plasmid vectors are used as transcription templates. As transcription proceeds to the end of DNA templates, plasmid linearization ensures that RNA transcripts of a defined length and sequence are generated. Yeasen provides various restriction enzymes to meet your needs. We recommend selecting restriction enzymes that generate blunt ends or 5´-overhangs. 

4.2 In Vitro Transcription ( IVT )

T7 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T7 phage promoters. In a cell-free system with DNA templates and NTPs, T7 RNA Polymerase catalyzes transcription from a cloned DNA sequence under the T7 promoters to synthesize RNAs. 

4.3 mRNA Capping

As a 5´ cap structure reduces immunogenicity and is required for efficient translation of mRNAs, adding cap structures to the 5′ ends of mRNA generated by transcription is necessary. 

Figure 1. Cap rate detection results (LC-MS detection)

4.4 mRNA Purification

Purification is an indispensable step to achieving high-purity mRNAs.

 

5. Raw materials for mRNA vaccine preparation provided by Yeasen

At present, Yeasen can provide a complete set of products in the production process of mRNA stock solution. The products are strictly controlled by GMP production and fully meet the requirements of mRNA vaccine production.

Table 2. Raw materials for mRNA vaccine preparation provided by Yeasen

Template preparation

In vitro transcription

Capping

Purification

Hieff Canace® Plus High-Fidelity DNA Polymerase

10153ES

Hifair® T7 High Yield RNA Synthesis Kit

10623ES

Vaccinia Capping Enzyme

10614ES

Hieff NGS® mRNA Isolation Master Kit

12603ES

dNTP Set Solution

10122ES

T7 RNA Polymerase

10624ES  10625ES

mRNA Cap
2´-O-Methyltransferase

10612ES

Hieff NGS® RNA Cleaner

12602ES

Hieff Clone® Plus One Step Cloning Kit

10911ES

Pyrophosphatase,Inorganic

10620ES

S-adenosylmethionine

10619ES

metal magnetic stand

80461ES

Hieff® Gold T4 DNA Ligase

10300ES

Murine RNase inhibitor

10621ES

Alkaline Phosphatase

Inquiry

 

 

Bsa I、BspQ I、Xba I

10661ES   10664ES  10662ES

DNase I

10611ES

CAP Analog

Inquiry

 

 

MolPure® Endo-free Plasmid Maxi Kit

19036ES

NTP Set Solution

10133ES

 

 

 

 

 

 

Pseudo UTP sodium solution GMP-grade (100 mM)

10650ES

 

 

 

 

 

 

N1-Me-Pseudo UTP sodium solution GMP-grade (100 mM)

10651ES

 

 

 

 

 

6. Ordering Information

The following are representative products offered by Yeasen. Additional sizes are available. Our products are highly optimized to work in concert, to help ensure superior performance and reproducibility. We can also provide customized services. If you’re interested in a product that isn’t shown, contact us and we’ll work with you to meet your needs.

Table 3. Ordering Information

Product name SKU Specifications
T7 High Yield RNA Synthesis Kit 10623ES 50/100/500 T
T7 RNA Polymerase GMP-grade (50 U/μL) 10624ES 5000/50000 U
T7 RNA Polymerase GMP-grade(250 U/μL) 10625ES 10/100 KU
10×Transcription Buffer GMP-grade 10627ES 1/10 mL
Pyrophosphatase,Inorganic GMP-grade 10620ES 10/100/1000 U
Murine RNase inhibitor GMP-grade 10621ES 10/20/100 KU
BspQI GMP-grade 10664ES 500/2500 U
DNase I GMP-grade 10611ES 500/2000/10000 U
mRNA Vaccinia Capping Enzyme GMP-grade 10614ES 2000/10000/100000 U
mRNA Cap 2'-O-Methyltransferase GMP-grade 10612ES 2000/10000/50000 U
10×Capping buffer GMP-grade 10666ES 1/10 mL
S-adenosylmethionine (SAM)(32 mM) 10619ES 0.5/25/500 mL
Pseudouridine-5-triphosphate,trisodium salt solution (100 mM) 10650ES 20 μL/100 μL/1 mL
N1-Me-Pseudo UTP sodium solution(100 mM) 10651ES 20 μL/100 μL/1 mL
ATP Solution(100 mM) 10129ES 1/25/500 mL
CTP Solution(100 mM) 10130ES 1/25/500 mL
UTP Solution(100 mM) 10131ES 1/25/500 mL
GTP Solution(100 mM) 10132ES 1/25/500 mL
NTP Set Solution (ATP, CTP, UTP, GTP, 100 mM each) 10133ES 1 Set (4 vial)
RNA Cleaner 12602ES 1/5/60/450 mL
mRNA Isolation Master Kit (Inquire) 12603ES 24/96 T
RNase H (Inquire) 12906ES 100/500/5000 U

 

Regarding reading:

GMP-grade reagents for mRNA in vitro synthesis

DNase I and Their Applications in Biomedicine

 

References:

[1] Chaudhary, N., Weissman, D., & Whitehead, K. A. (2021). mRNA vaccines for infectious diseases: principles, delivery and clinical translation. Nature reviews. Drug discovery, 20(11), 817–838.
[2] Vogel, A. B. et al. Self-Amplifying RNA Vaccines Give Equivalent Protection against Inflfluenza to mRNA Vaccines but 
at Much Lower Doses. Molecular therapy : the journal of the American Society of Gene Therapy 26, 446-455, 
[3] Fuchs, A. L., Neu, A. & Sprangers, R. A general method for rapid and cost-effiffifficient large-scale production of 5' 
capped RNA. RNA (New York, N.Y.) 22, 1454-1466, doi:10.1261/rna.056614.116 (2016). 
[4] Schmid, A. Considerations for Producing mRNA Vaccines for Clinical Trials. Methods in molecular biology (Clifton, N.J.) 1499, 237-251, 
[5] Banerji, A. et al. mRNA Vaccines to Prevent COVID-19 Disease and Reported Allergic Reactions: Current Evidence and Suggested Approach. The journal of allergy and clinical immunology. In practice 9, 1423-1437, 
[6] Richner, J. M. et al. Modifified mRNA Vaccines Protect against Zika Virus Infection. Cell 168, 1114-1125.e1110, 
[7] Corbett, K. S. et al. SARS-CoV-2 mRNA vaccine design enabled by prototype pathogen preparedness. Nature 586,567-571, 
[8] Corbett, K. S. et al. mRNA-1273 protects against SARS-CoV-2 beta infection in nonhuman primates. Nature immunology 22, 1306-1315, 
[9] Keech, C. et al. Phase 1-2 Trial of a SARS-CoV-2 Recombinant Spike Protein Nanoparticle Vaccine. The New England journal of medicine 383, 2320-2332,
[10] Kramps, T. & Elbers, K. Introduction to RNA Vaccines. Methods in molecular biology (Clifton, N.J.) 1499, 1-11, 
[11] Linares-Fernández, S., Lacroix, C., Exposito, J. Y. & Verrier, B. Tailoring mRNA Vaccine to Balance Innate/- Adaptive Immune Response. Trends in molecular medicine 26, 311-323,