mRNA Quality Control (QC) is a foundational analytical process to verify therapeutic transcripts meet specs for identity, purity, potency, and safety—without it, minor impurities (e.g., fragmented transcripts, dsRNA) harm efficacy, trigger adverse immunity, and halt clinical development.
Creative Biolabs' End-to-End mRNA QC Platform addresses translation bottlenecks, contaminant immunogenicity, and regulatory challenges via advanced analytics, streamlining biomanufacturing for high-purity, stable transcripts. We specialize in comprehensive Critical Quality Attribute (CQA) assessment, ensuring chemical purity, optimized in vivo translation, and minimized innate immune activation to bridge synthesized mRNA to safe, effective therapeutics.
Discover How We Can Help - Request a Consultation TodaymRNA QC requires comprehensive verification of attributes "from molecular structure to biological function," with a core focus on 6 key dimensions to ensure the product meets R&D or clinical application requirements:
Verify synthesized mRNA matches the target sequence to avoid sequence error-caused functional failure or safety risks. Confirm mRNA's nucleotide sequence and key modifications (e.g., 5' cap, 3' Poly(A) tail, modified nucleotides) align with the design.
Remove synthesis-generated impurities (e.g., template DNA, unreacted NTPs, dsRNA, truncated fragments) that may trigger immunogenicity (e.g., dsRNA activating innate immunity) or reduce translation efficiency.
Confirm that mRNA molecules are free from degradation or breakage, and that their molecular weight matches the theoretical value—degraded or broken mRNA loses translational function and cannot exert therapeutic effects.
Accurately determine mRNA concentration to ensure controllable dosage in subsequent experiments or formulation production; simultaneously verify the homogeneity of mRNA molecules within a batch (no fragments with significant size differences).
For mRNA containing chemical modifications (e.g., pseudouridine ψ, N1-methylpseudouridine m1ψ), verify the incorporation ratio and site accuracy of modified nucleotides—modification efficiency directly affects mRNA's immunogenicity (reducing innate immune activation) and stability.
Verify mRNA's translational ability in vitro/in vivo—only mRNA that can effectively express the target protein can meet vaccine needs (stimulating immune responses) or therapeutic needs (supplementing functional proteins).
| Testing Dimension | Common Technical Methods | Core Function |
|---|---|---|
| Identity | RT-PCR, Next-Generation Sequencing (NGS), Mass Spectrometry | RT-PCR/NGS verifies nucleotide sequences; MS confirms 5' cap structures (e.g., Cap 1) and types of modified nucleotides |
| Purity | Anion Exchange High-Performance Liquid Chromatography (AEX-HPLC), Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC), DNase Digestion Assay | AEX-HPLC removes dsRNA/truncated fragments; RP-HPLC removes proteins/salts; DNase digestion assay detects residual DNA |
| Integrity & Molecular Weight | Capillary Electrophoresis, Agarose Gel Electrophoresis (CE/AGE), Mass Spectrometry (MS) | CE/AGE visually shows whether mRNA is degraded (no heterobands); MS measures molecular weight for comparison with theoretical values |
| Concentration & Homogeneity | Ultraviolet-Visible Spectrophotometer (UV-Vis), Fluorometric Quantitation Kits | UV-Vis quantifies concentration via A260/A280 ratio (ideal range: 1.8-2.0); RiboGreen improves detection sensitivity for low-concentration mRNA |
| Modification Efficiency | Liquid Chromatography-Tandem Mass Spectrometry (LC-MS), High-Performance Capillary Electrophoresis (HPCE) | LC-MS/MS accurately quantifies the incorporation rate of modified nucleotides; HPCE separates modified and unmodified mRNA to evaluate modification uniformity |
| Functional Activity | Cell Transfection Assay and Western Blot/ELISA, Reporter Gene Assay | Western Blot detects target proteins after transfection; reporter genes (e.g., Luciferase) rapidly quantify translation efficiency |
Our process is designed for maximum clarity, reproducibility, and efficiency, providing traceable results suitable for regulatory submission.
| Workflow Step | Activities Involved |
|---|---|
| Project Scoping & CQA Definition | Initial consultation to define project goals (e.g., preclinical, Phase I), source material, and agree on Critical Quality Attributes (CQAs) and acceptance criteria. |
| Integrity & Purity Analysis | Use Capillary Electrophoresis (CE) and high-resolution gel analysis to determine mRNA length, fragmentation, and purity (vs. contaminants like truncated transcripts). |
| Critical Structure Verification | Apply advanced LC-MS/MS or validated enzymatic assays to quantify 5' cap efficiency (Cap 0/Cap 1) and confirm poly(A) tail length/fidelity (key for stability). |
| Contaminant & Safety Testing | Run ultra-sensitive qPCR for residual template DNA, and specialized ELISA/immune-activation assays for dsRNA and host cell protein/reagent remnants. |
| Final Data Package & Release | Compile analytical data, raw files, Certificate of Analysis (CoA), final report, and provide expert consultation on interpretation. |
Required Starting Materials:
Final Deliverables:
Estimated Timeframe: The typical timeframe for this service ranges from 2 to 4 weeks, depending on the number of assays required and the complexity of the client's material (e.g., novel capping strategy, non-standard nucleoside modification).
At Creative Biolabs, we recognize that every mRNA therapeutic is unique, demanding a QC strategy that is precise, comprehensive, and tailored to its specific Critical Quality Attributes. As your dedicated analytical partner, we offer flexible, high-resolution services designed to move your construct swiftly and safely toward the clinic.
Targeted Cap Structure Quantification
Precise LC-MS/MS and HPLC methods to accurately verify 5' Cap integrity (Cap 1 vs. Cap 0), directly correlating analytical results with translational potency.
Ultra-Sensitive dsRNA and DNA Clearance
Specialized qPCR and affinity-based assays to quantify highly immunogenic double-stranded RNA and residual DNA templates, ensuring safety thresholds are met for regulatory submission.
Full-Length Integrity Analysis (FLI)
High-resolution Capillary Electrophoresis (CE) and gel-based fractionation to confirm the stability and complete FLI of long and complex mRNA constructs up to 10 kb.
Customized Assay Development and Validation
Creative Biolabs designs, optimizes, and validates bespoke analytical methods to qualify mRNA constructs utilizing novel chemistries, modified nucleosides, or non-canonical UTRs tailored specifically to your research needs.
Seamless Process Integration
Dedicated project management and scientific consultancy to ensure our QC data integrates smoothly with your IVT and purification workflows, minimizing process bottlenecks.
A: Even highly purified products can harbor trace but potent immunogenic impurities like dsRNA or residual DNA, which are difficult to remove and detect using standard methods. Our ultra-sensitive qPCR and specialized assays target these specific contaminants to ensure your product meets the required safety profile for in vivo use.
A: All Creative Biolabs data packages are generated using validated methods with full audit trails, making them compatible with cGMP principles. We provide expert consultation to help interpret the data and ensure it aligns perfectly with the requirements for IND or BLA submissions.
A: Absolutely. We utilize high-resolution electrophoretic and specialized analytical techniques designed to handle large transcripts up to 10 kb. The key to a functional therapeutic is non-fragmented, full-length integrity, and our robust methodology confirms the structural fidelity of even the longest mRNA constructs.
Creative Biolabs is your trusted partner for therapeutic mRNA development. We offer validated, comprehensive, and compliance-driven Quality Control solutions that extend beyond standard analysis to guarantee the functional integrity and safety of your transcripts. Leverage our expertise to minimize risk, maximize translational efficacy, and accelerate your path from discovery to clinic.
Contact Our Team for More Information and to Discuss Your Project