mRNA & LNP Therapeutics

mRNA & LNP Therapeutics (IVT → LNP → Fill/Finish)

Mika Biologics — your end-to-end mRNA CDMO

Why this service exists

If you’re developing mRNA medicines, you already know that success is not one step—it’s a chain. High-quality plasmid DNA, clean IVT chemistry, decisive dsRNA reduction, robust purification, and a reproducible LNP that holds size, PDI, and potency under real-world handling. Most programs stumble because one link isn’t manufacturable. Mika Biologics built this service to close the loop.

Use cases we support

  • Infectious disease vaccines: IM dosing with stability at 2–8 °C or frozen; rapid pivot for new variants.
  • Oncology (cancer vaccines & cytokines): Personalized mRNA, shared-neoantigen targets, immune stimulants with controlled innate activation.
  • Protein replacement / gene regulation: mRNA encoding secreted or intracellular proteins; exon-skipping strategies; gene editing payloads requiring precise integrity and delivery.
  • Ex vivo cell engineering: mRNA for CAR-T/NK editing and transient expression, where clean IVT and low dsRNA reduce cytotoxicity and innate sensing.
  • Platform enablement: Method transfers, process characterization, comparability runs, bridging to pivotal.

Every use case is governed by a clear QTPP → CQAs → CPPs map (below) so design choices stay tied to clinical goals.

Process flow overview (pDNA → IVT → Purification → LNP → DP)

Plasmid DNA upstream (GMP or high-quality R&D)

  • Template design: ORF, UTRs, poly(A) tail strategy, potential N1-methyl-pseudouridine substitution, and sequence-level dsRNA risk.
  • Linearization: High-fidelity restriction digest or enzymatic approach with orthogonal confirmation of complete linearization.
  • Quality inputs: Residual host DNA, RNA, protein, and endotoxin specifications to prevent carryover.
    Internal link: See our GMP Plasmid DNA page for upstream supply continuity.

IVT mRNA manufacturing

  • Reaction: Linearized pDNA → IVT using T7 RNAP; Mg²⁺, rNTP ratios, and reaction temperature/time tuned by DoE.
  • Capping options:
    • Co-transcriptional: ARCA, CleanCap, or other cap analogs;
    • Enzymatic: post-transcriptional capping with high cap-1 efficiency.
  • Poly(A) strategy: Tailed template or enzymatic polyadenylation with controlled distribution.
  • Template removal: DNase digestion with validated clearance.
  • dsRNA reduction: Process design (nucleotide balance, temperature, cap chemistry) plus downstream HPLC/TFF and cellulose-based or chromatographic dsRNA removal steps.
  • Primary polish: Tangential flow filtration, anion-exchange, or reversed-phase/HIC options depending on molecule and program phase.

Purity & integrity confirmation

  • Integrity: Bioanalyzer/Fragment Analyzer, CGE, or agarose to confirm full-length.
  • Identity: Sequence confirmation (NGS/Sanger), restriction mapping, or enzymatic fingerprinting.
  • Capping efficiency: LC-MS or enzymatic assay.
  • Poly(A): Tail length profiling.
  • dsRNA: ELISA/dot-blot (J2 antibody) with acceptance criteria set from QTPP.
    Internal link: Deeper methods on our Analytical & QC page.

LNP formulation (design → data → decision)

  • Lipid system: Ionizable lipid + helper phospholipid + cholesterol + PEG-lipid tuned for route (IM/IV), dose, and stability.
  • Microfluidic mixing: FRR (aqueous:organic), TFR, and lipid:mRNA ratio optimized; scalable mixers from discovery to GMP.
  • DoE grid: N/P ratio, % ionizable lipid, PEG chain length & mol%, buffer pH/ionic strength, mixing parameters.
  • Targets: Size 60–100 nm (program-dependent), PDI ≤ 0.2, encapsulation efficiency ≥ 85–95%, narrow distribution, low ethanol residuals.
  • Diafiltration / buffer exchange: Ethanol removal below ICH Q3C limits; osmolality and pH set for administration route.

Drug Substance & Drug Product

  • DS (bulk LNP or mRNA): Held at qualified temperatures; stability-indicating assay panel defined.
  • DP (final vials): Sterile filtration (0.22 µm where appropriate), aseptic fill-finish, labeling, and pack-out for frozen or refrigerated supply chains.
    Internal link: See Formulation & Aseptic Fill-Finish for line configuration and components.

What we deliver (scope recap)

  • IVT mRNA: linearized pDNA → IVT → DNase → co-transcriptional or enzymatic cap (ARCA/CleanCap/enzymatic) → poly(A) → HPLC/TFF polish with dsRNA reduction.
  • LNP formulation: Ionizable lipid selection, DoE on N/P, size/PDI tuning, encapsulation efficiency optimization.
  • Analytical/QC: Integrity, capping %, dsRNA ELISA, potency, residuals (DNA, protein, solvents), endotoxin, DLS, LC-MS.
  • DS/DP: Sterile filtration, frozen storage at −20 °C/−80 °C, tech transfer, CMC authoring for IND/IMPD.

QTPP → CQA → CPP: how we design for manufacturability

Quality Target Product Profile (QTPP) (examples)

  • Route (IM/IV), dose per vial, storage condition (−20 °C/−80 °C, or 2–8 °C where justified)
  • Onset of action and durability targets
  • Safety/tolerability: controlled innate activation, low endotoxin, ethanol residuals below limits
  • Presentation: vial fill volume, concentration, container-closure system

Critical Quality Attributes (CQAs)

  • mRNA: identity, full-length integrity, purity, capping efficiency, poly(A) length, dsRNA content, residual DNA/protein/solvents, endotoxin, bioburden/sterility for DP
  • LNP: size, PDI, encapsulation efficiency, pH/osmolality, zeta potential (if applicable), lipid composition, residual ethanol, appearance, potency
  • Stability: real-time/accelerated profiles, freeze–thaw robustness, shipping vibration/handling tolerance

Critical Process Parameters (CPPs)

  • IVT: temperature, time, Mg²⁺, NTP ratios, cap analog stoichiometry, template concentration, DNase conditions
  • Polish: TFF TMP & shear, membrane MWCO, chromatography gradients, dsRNA removal load/flow
  • LNP: FRR/TFR, N/P ratio, lipid:mRNA ratio, ethanol %, buffer pH/ionic strength, mixing geometry
  • Fill-finish: filtration differential pressure, hold times, fill accuracy, stopper venting, headspace, lyophilization (if used)

By fixing CQAs that matter to the clinic and mapping CPPs that move those CQAs, our mRNA CDMO approach avoids “pretty data” that fail in scale-up.

Analytics panel (expanded)

Identity & integrity

  • CGE/Fragment Analyzer/Bioanalyzer: full-length confirmation, size distribution
  • NGS or Sanger for template identity; restriction fingerprinting as needed

Cap & poly(A)

  • Cap efficiency: LC-MS or enzymatic assays (e.g., cap-specific detection)
  • Poly(A) length: electrophoresis or sequencing-based profiling

Purity & dsRNA

  • HPLC variants (AEX/RP/HIC) to quantify purity and product-related variants
  • dsRNA quantitation: J2 ELISA or dot-blot; acceptance criteria set per indication and route

Residuals

  • Template DNA, RNase, protein, enzymes (e.g., T7): qPCR, ELISAs
  • Ethanol & other solvents: GC/HS-GC against ICH Q3C limits
  • Endotoxin & bioburden/sterility: per USP/EP where applicable

LNP characterization

  • DLS: size and PDI; number-weighted where useful
  • Encapsulation efficiency: RiboGreen or equivalent dye-exclusion methods
  • Composition confirmation: TLC/LC methods for lipid ratios; optional zeta potential

Potency

  • In vitro expression assays (e.g., luciferase or target protein) in relevant cell lines
  • Innate activation profiling (as requested): cytokine readouts after transfection to benchmark dsRNA reduction success

Stability

  • Accelerated and real-time across target storage; freeze–thaw cycling; agitation/shipping studies
  • Container–closure compatibility and extractables/leachables strategy

Internal links: More detail on our Analytical & QC capabilities. If you require bulk T7 RNAP or assay enzymes, see Diagnostics & IVD Enzymes.

Mika Biologics, CDMO for Microbial projects

LNP formulation strategy: design space that holds in the real world

Ionizable lipid selection: We start with your preferred family or propose candidates aligned to pKa and route of administration.
Helper lipids & PEG-lipid: Cholesterol % for membrane stability; helper phospholipid headgroup for bilayer behavior; PEG density tuned to circulation and aggregation risk.
DoE blocks:

  • N/P ratio sweep to balance encapsulation and cytotoxicity
  • FRR/TFR to drive nucleation/size; scale-aware parameters that translate from bench to GMP mixers
  • Buffer & pH to set ionization and storage behavior
    Acceptance criteria (program-specific): typical size 60–100 nm, PDI ≤ 0.2, high encapsulation, low ethanol.

We don’t “lock” a single recipe too early; instead, we fix a range that’s robust to supplier change, temperature fluctuations, and real cold chain handling. That’s how a LNP CDMO protects your CMC risk.

Scale & timelines (typical ranges)

Development scales (non-GMP):

  • IVT: milligrams → multi-grams per batch for screening and analytics qualification
  • LNP: milliliters → multi-liters for DoE, stability, and early tox lots

GMP scales:

  • IVT: multi-gram to tens-of-grams per batch, program dependent
  • LNP DP: clinical lots configured to dose & vial strategy (single- or multi-dose as appropriate)

Illustrative timeline archetype (subject to scope):

  1. Feasibility (4–6 weeks): template prep/transfer, IVT screen, dsRNA baseline, initial LNP feasibility
  2. Process development (8–12 weeks): DoE for IVT and LNP, dsRNA reduction locking, stability-indicating methods
  3. GMP readiness (6–10 weeks): documentation, method qualification, engineering runs, PQs as required
  4. GMP manufacture (lot-specific): DS/DP production, QC release, quality review, batch record issuance

Timelines vary with sequence complexity, analytics scope, and regulatory strategy. Ask us to tailor a Gantt to your program.

Regulatory/CMC package for IND/IMPD

Documentation set (indicative):

  • Process description: narrative + flow diagrams for IVT, purification, LNP, fill-finish
  • Materials & suppliers: specs/COAs, qualifications, change-control alignment
  • Master/Batch records: DS and DP with hold times and IPCs
  • Analytical methods & validation/qualification status
  • Specifications & justifications: CQAs and limits tied to clinical route and risk assessments
  • Stability protocols & data: real-time/accelerated; shelf life rationale
  • Comparability plans: pre-pivotal/pivotal transitions, formulation/scale/source changes
  • Risk assessments: including dsRNA risk and endotoxin/LER strategies
  • Cleaning validation / cross-contamination controls for shared equipment where applicable

We work to the regional dossier structure (e.g., IND Module 3 / IMPD sections) and coordinate with your regulatory team or external agents for alignment.

Tech transfer & risk controls

Upstream tech transfer

  • Sequence & plasmid package; SOPs; historical batch data; past deviations/lessons learned
  • Bilateral method gap assessment; material compatibility studies (buffers, membranes)

Manufacturing risk controls

  • RNase control: facility zoning, single-use flow, trained gowning, RNase-free consumables, activity checks
  • Particulate control: filtration strategy plus environmental monitoring and visual inspection
  • Endotoxin control: low-bioburden water and materials, recovery checks, and LER-aware validation
  • Solvent control: ethanol removal validated by GC; flammability/EHS procedures documented
  • Chain of custody: full traceability from raw materials to final vials

Knowledge transfer

  • Full document pack; equipment lists and set-points; deviation history; scale-down models for investigations
  • Joint PPQ design where relevant before pivotal manufacture

Manufacturing options & add-ons

  • Modified nucleosides: N1-methyl-pseudouridine, 5-methyl-CTP, etc., with supplier qualification
  • Sequence optimization support: codon optimization, UTR tuning (optional, discovery stage)
  • Lyophilization feasibility: where route/formulation justify; cake and reconstitution studies
  • Alternate delivery systems: polymeric nanoparticles or other non-LNP carriers (case-by-case)
  • Cold chain packaging: validated shippers for −20 °C/−80 °C; 2–8 °C stability if supported
  • Reference standard generation and reserve samples strategy

What makes Mika different as your mRNA CDMO

  • Manufacturability by default: We start every program with a QTPP→CQA→CPP table that governs both science and documentation.
  • Real DoE, not trial-and-error: Formal experimental designs for IVT and LNP—so the parameters you qualify are the ones you can defend.
  • Analytical depth: From cap analysis and dsRNA ELISA to lipid composition checks and in-vitro potency—bench to release without “method drift.”
  • Single-partner continuity: pDNA → IVT → LNP → Fill-Finish under one quality system.
  • Change-proofing: Alternative vendors and acceptable ranges pre-qualified to cushion supply volatility.

FAQ (mRNA CDMO & LNP CDMO)

1) Do you require us to use your plasmid DNA?
No. We can start with your qualified template or supply GMP Plasmid DNA from our network. We do verify linearization and quality before IVT.

2) What’s your typical dsRNA acceptance range?
It’s indication- and route-specific. We set a specification based on your QTPP and demonstrate process capability using dsRNA ELISA/dot-blot. We can drive dsRNA down with process and polish optimizations.

3) Can you hit a specific LNP size and PDI?
Yes—within a process window established by DoE (FRR/TFR/N/P/buffer). Many clinical programs target 60–100 nm with PDI ≤ 0.2; we’ll align targets to your route and dose.

4) Do you support enzymatic capping and cap analysis?
Yes. We run co-transcriptional or enzymatic capping and quantify capping % using LC-MS or enzymatic methods.

5) How do you control endotoxin and solvent residuals?
Low-endotoxin materials and rigorous environmental controls, with GC for ethanol residuals against ICH limits and USP methods for endotoxin/bioburden/sterility.

6) What stability can I expect?
Program-dependent. We develop stability-indicating assays, run accelerated/real-time studies, and qualify storage at −20 °C/−80 °C (or 2–8 °C where supported).

7) Can you do potency testing?
Yes—standard in-vitro expression assays in relevant cells. For custom pharmacology, we collaborate with your biology team or partner CROs.

8) How do you handle tech transfer from my current CDMO?
Documented transfer with side-by-side method checks, engineering runs, and a comparability plan. We preserve your prior learnings and lock new process parameters with data.

9) Do you support rapid variant updates for vaccines?
Yes. With template changes, we execute a risk-based comparability plan and leverage established CPP windows for fast-track production.

10) Do you provide LNP alternatives?
We can explore polymers or hybrid systems as a development track; for clinical timelines, LNP remains the fastest route to a qualified CMC package.

How to engage

  1. Discovery call: Align on QTPP, clinical route, dose, and timelines.
  2. Data review: Your sequence and historical data; our proposed DoE and analytics plan.
  3. Workplan & quote: Clear milestones, deliverables, and decision gates.
  4. Kickoff: Materials list, schedule, and risk register.

Internal links:

  • GMP Plasmid DNA — continuous upstream supply and documentation
  • Analytical & QC — method depth, validation/qualification plan
  • Formulation & Aseptic Fill-Finish — vialing lines, components, sterility strategy
  • Diagnostics & IVD Enzymes — if you require T7 RNAP or auxiliary enzymes under quality control

Summary

Your mRNA program needs more than a vendor—it needs a mRNA CDMO that designs for manufacturability from day one. Mika Biologics integrates IVT, dsRNA reduction, HPLC/TFF purification, LNP formulation, analytics, and sterile fill-finish into a single, defensible process built on QbD. The outcome is simple: a data-backed CMC narrative you can take to regulators and a product that behaves in the clinic and in the real world.

Let’s build your process—once.

Email our sales team at info@mikabiologics.com