mRNA has been widely used in the study of gene or protein functions. Meanwhile, it is common in the development of prophylactic and therapeutic biologics. As a common synthesis strategy for RNA molecules, in vitro transcription (IVT) enables us to obtain customized transcripts efficiently.
Yaohai Bio-Pharma has established a set of mRNA synthesis technologies to provide high-quality RNA, such as sequence design and optimization, IVT, purification, lyophilization and lipid nanoparticle (LNP) encapsulation. All products are released under stringent quality control (QC) standards.
With the outbreak of the COVID-19 pandemic in 2020, mRNA technology has advanced by leaps and bounds, and its related research and applications have gained unprecedented development, including prophylactic vaccines for infectious disease prevention, therapeutic oncology vaccines and drugs, tumor therapy, protein replacement therapy, regenerative medicine, and cell and gene therapy (CGT).
mRNA Vaccine | mRNA Therapeutics |
|
Process | Optional Service | Service Details | Delivery Period (workday) | Deliverable |
mRNA Sequence Design and Optimization | Design and optimization of coding sequences | CDS sequence design and codon optimization | 1-3 | Sequence file |
Design and optimization of non-coding sequences | Design and optimization of UTR, polyA sequences | |||
Transcription Template Plasmid Preparation | Recombinant plasmid preparation | Gene synthesis | 7-10 | |
Plasmid amplification and extraction | 4 | |||
Plasmid linearization and purification | ||||
mRNA In Vitro Transcription | Co-transcriptional capping (one-step method) | In vitro transcription (IVT) / Cap analog | 1-2 | N/A |
Nucleotide modifications (UTP/CTP modifications) | ||||
DNA template removal (DNase I) | ||||
Enzymatic capping (two-step process) | IVT | 2-3 | ||
Nucleotide modifications (UTP/CTP modifications) | ||||
DNA template removal (DNase I) | ||||
mRNA purification (lithium chloride/magnetic beads) | ||||
Enzymatic capping | ||||
mRNA Purification | Conventional purification scheme | Lithium chloride precipitation | 1 | mRNA drug substance |
Magnetic bead purification | ||||
Chromatography column purification scheme | Combination of multiple chromatography methods | 1-2 | ||
Buffer exchange | Ultrafiltration | 1 | ||
mRNA Lyophilization | Lyophilization | Pre-freezing | 2-3 | mRNA lyophilized powder |
Primary drying (sublimation) | ||||
Secondary drying (desorption) | ||||
mRNAEncapsulation | LNP encapsulation | LNP encapsulation | 2-3 | mRNA-LNPDrug product |
Concentration and buffer exchange | ||||
mRNA Quality Analysis | mRNA drug substance/ lyophilized powder | Concentration, purity | 1 | Test Report |
Integrity, capping efficiency, polyA tail distribution | 2-5 | |||
mRNA-LNP drug product | Encapsulation efficiency | 1 | ||
Particle size and distribution | ||||
Surface charge | ||||
mRNA Expression Validation | 293T cell evaluation | Cell plating | 4 | |
Transient transfection of cells | ||||
Fluorescence signal observation | 1-3 | |||
Western blot (WB)/ Enzyme-linked immunosorbent assay (ELISA) |
5' Cap |
▪ Uncapped ▪ Cap0, co-capped ▪ Cap1, co-capped ▪ Cap1, enzymatic capping |
5’ UTR/3’ UTR |
▪ Nature UTR sequence ▪ Mutant/Engineered UTR sequence |
3' PolyA Tail |
▪ 100A ~120A Tail (recommended) ▪ Segmented polyA tail ▪ Other Custom tails |
Modified nucleosides |
▪ Unmodified bases ▪ Pseudouridine (Ψ) ▪ N1-Methylpseudouridine (N1Ψ) ▪ 5-methylcytosine (m5C) ▪ 5-methyluridine (m5U) ▪ 5-methoxyuridine (5moU) ▪ 2-thiouridine (s2U) ▪ 2′-O-methyl-U ▪ Other modifications |
Integrated Service Flow
Provide a series of services from front-end sequence design to back-end mRNA preparation, quality control and expression validation.
International Cutting-edge Sequence Design and Optimization
Professional mRNA sequence design and optimization facilitate efficient mRNA expression.
Diversified Nucleotide Modifications
Effectively increase mRNA expression and reduce mRNA adverse immune responses.
Mature Purification Platform
By combining a conventional and self-designed purification method, mRNA samples are obtained with high purity
Complete QC Platform
Enrich QC options to meet the requirements of routine tests, such as concentration, A260/280 purity, and integrity, as well as high quality control requirements, such as capping efficiency/polyA distribution.
Fast Delivery
Customized mRNA can be delivered in as fast as 7 days except for outsourced sequence synthesis.