FDA has Approved Yeasen’s Master Files

With the rapid development of biotechnology, mRNA vaccines produced by in vitro RNA

synthesis technology have had a huge impact on the outbreak of COVID-19 in 2020. Many

companies, such as Moderna(US), CureVac(Germany), and BioNTech, have already

developed therapeutic mRNA vaccines and drugs for other diseases such as tumours,

infectious diseases, and chronic diseases.

Figure 1. Schematic diagram of mRNA vaccine and its antigen expression mechanism [1]

The mRNA vaccine development process includes pathogen identification, sequencing,

mRNA synthesis and modification in vitro, and purification and so forth. The development

and production of mRNA vaccines require a variety of raw materials to ensure the successof vaccines. As an innovative leader in the domestic molecular enzyme industry, Yeasen

Biotechnology has successfully developed various technologies through advanced

molecular enzyme two-way technology platforms and large-scale protein fermentation and

purification technology platforms. Products of similar molecular enzyme-related raw

materials. Yeasen Biotechnology provides GMP grade raw materials required for mRNA

vaccine research and is recognized by a large scale of customers.

Difficulties in preparing mRNA vaccines

The production of mRNA vaccines is mainly through in vitro synthesis (IVT). The process is

mainly based on DNA as a template plus corresponding substrates and buffers through in

vitro transcription. Because the stability of RNA is weak, the stability of mRNA after

synthesis is a problem that must be paid attention to. The market we face is that enzyme

raw materials are mainly imported, which means the international supply chain can be cut

at any time. At the same time, patent barriers also restrict the development of the industry

in terms of capping analogues. In addition, the quality of various domestic raw material

manufacturers is unstable. The lack of universal industry standards has also become an

obstacle to the development of domestic upstream and downstream enterprises.

Figure 2. Difficulties in mRNA preparationRaw Material for mRNA Production

After ingenious research and development, Yeasen Biotechnology provides a complete set

of raw materials around the mRNA production process, covering various enzymes and

substrates. Gradually reduce import dependence. The products are strictly controlled by

GMP standard, which fully meets the requirements for mRNA vaccine production.

Figure 3. The raw materials for the production of mRNA

Our products have corresponding product solutions in each step from in vitro transcription

of mRNA preparation to mRNA modification and purification to meet the different needs of

customers.

Figure 4. Yeasen provides raw materials for in vitro synthesis of GMP-grade mRNA

mRNA in Vi t ro Tr ans c r ipt ion

mRNA Modification

mRNA Purification

IVT

Professional solutions from template, transcription to modification and purificationThe cost-effective Hifair® T7 High Yield RNA Synthesis Kit developed by Yeasen contains a

complete set of reagents required for RNA synthesis and a detailed tutorial.

Product Features

High yield: 100-200 µg can be produced within 2 hours, and a single amplification reaction

can produce mg-level RNA

Multiple scenarios: suitable for long and short RNA transcription

High flexibility: get unlabeled, labelled and capped RNA

Short fragment: 2-4 h can be effectively transcribed

Long fragment: 1-2 h can be effectively transcribed

Figure 5. Transcription yields of different target fragments

mRNA modification

Yeasen provides low-residue and high-efficiency Vaccinia Capping Enzyme by adding a cap

structure to the 5'end to enhance the stability of mRNA.

Product Features

High standard: GMP grade production

No residue: no nuclease, low endotoxin content

High purity: purity >95% (SDS-PAGE)

High specific activity: ~160000 U/mg (BCA)

Capping efficiency

Measure the capping efficiency of the enzyme by mass spectrometry and compare it withthe reference. It can be found that the capping efficiency of Yeasen products is relatively

high. Cap1 capping can reach 100%, and the purity of the obtained product is also high.

Figure 6. Enzymatic capping efficiency: control group (left) and experimental group

Cap1 (right)

Yeasen Biotechnology GMP Grade mRNA in Vitro Synthesis

Raw Material Production Capacity

In terms of solving the shortage of raw materials, Yeasen has sufficient production

capacity and has built an ultra-clean factory of molecular enzyme (UCF.ME) covering an

area of 3000 square meters and a fermentation scale of 800 L per patch in Wuhan. Yeasen

is the first Molecular enzyme manufacturer which has obtained ISO 13485: 2016 quality

system certification in China.Figure 7. Yeasen mRNA raw material production capacity

Figure 8. Yeasen molecular enzyme products obtained ISO 13485: 2016 quality system

certificationFigure 9. A picture of Yeasen Ultra Clean Factory of Molecular Enzyme (UCF.ME)

production base

Establishing a complete quality management system to provide better quality products has

always been our direction, in addition to the current industrial-grade AKTA purification and

physical and chemical analysis equipment, 100L scale of high-density fermentation units,

clean workshops, and automated production lines of the ultra-clean factory of molecule

enzyme (UCF.ME),.To expand production that can meet a wider demand from customers,

we have put forward more stringent requirements and deployed a 4,500 square meter

mRNA Tools production base, which is specifically designed for the production of mRNA

vaccine raw materials.

·Higher quality management system:

Designed and constructed in accordance with the pharmaceutical production plant, fully

complying with pharmaceutical GMP standards

·The production capacity is improved:1500ully automatic fermentation system X 2;

·More abundant production lines:

GMP grade chemical synthesis workshop, producing NTP, SAM, capped analogues

mRNA Production Relative Products

Procedure

Product Name

NO

Size

IVT

Hifair® T7 High Yield RNA Synthesis Kit

10623ES50/60

50/100 T

UCF.ME® T7 RNA Polymerase GMP-grade（

50 U/μL） 10624ES86/90/96/97

2.5/5/25/50 KU

UCF.ME® T7 RNA Polymerase GMP-grade（1 KU/μL） 10613ES92/97/98/99

10/50/500/5000

KU

UCF.ME® Deoxyribonuclease I (DNase I) GMP-grade

10611ES76/84/92

0.5/2/10 KU

UCF.ME® Pyrophosphatase, Inorganic GMP-grade

10620ES10/60/80

10/100/1000 U

UCF.ME® Murine RNase inhibitor GMP-grade

10621ES10/20/60

10/20/100 KU

NTP Set Solution (ATP, CTP, UTP, GTP, 100 mM

each）

10133ES03

1 Set (4 vial)

mRNA

modification

UCF.ME® mRNA Vaccinia Capping Enzyme

GMP-grade

10614ES84/92/94/96

2/10/20/100 KU

UCF·ME® mRNA Cap 2´-O-Methyltransferase

GMP-grade

10612ES84/92/97/98

2/10/50/250 KU

S-adenosylmethionine (SAM)（

32mM）

10619ES02/25/76

0.5/25/500 mL

GTP (100 mM)

10132ES03

1 mL

mRNA

purification

Hieff NGS® RNA Cleaner

12602ES03/08/56/75

1/5/60/450 mL

【1】Verbeke R, Lentacker I, Smedt S, & Dewitte H (2019) Three decades of messenger

RNA vaccine development. Nano Today:100766.