| Field | Specification |
|---|---|
| Mfr No | |
| Activity | |
| Biological Activity | |
| Concentration | |
| Endotoxin Level | |
| Enzyme Type | |
| Expression System | |
| Form | Liquid (sterile filtered, ready-to-use) |
| Formulation | |
| Product Type | |
| Purity | |
| Source | Recombinant fungal enzyme (Fusarium oxysporum) produced via Bacillus sp. fermentation |
| Species | |
| Storage |
Scientific Background
Trypsin-like serine proteases catalyze peptide bond cleavage at the carboxyl side of lysine and arginine residues — the same enzymatic mechanism exploited during cell dissociation (trypsinization). Conventional porcine trypsin introduces biosafety concerns associated with animal-derived materials, including batch-to-batch variability and potential adventitious agent contamination. Recombinant, non-mammalian alternatives address these risks by enabling consistent, animal origin-free production under controlled manufacturing conditions.
CellTrypase is a recombinant trypsin-like enzyme derived from Fusarium oxysporum, produced via microbial fermentation using Bacillus sp. as the expression host. Its fungal origin confers inherently lower non-specific reactivity compared to mammalian trypsin, resulting in gentler cell dissociation with preserved viability and function.
Product Description
Expression System: Microbial Fermentation (Bacillus sp.)CellTrypase is manufactured in-house by c-LEcta GmbH (a Kerry Company) under a certified Quality Management System. It is available in two quality grades — GMP-grade (EXCiPACT® GMP/GDP certified) and R&D-grade (ISO 9001 compliant) — and in two concentration formats (1x ready-to-use; 10x concentrated stock), supplied in 100 mL and 500 mL bottle sizes.
The formulation is a sterile-filtered liquid in PBS with EDTA. By eliminating drying and reconstitution steps, the liquid format minimizes degradation risk and preserves enzyme integrity — yielding a crystal-clear solution that visibly reflects its purity.
Technical Specifications
| Enzyme Origin | Recombinant Fusarium oxysporum expressed in Bacillus sp. |
|---|---|
| Enzyme Class | Serine Protease (Trypsin-Like) |
| Cleavage Specificity | Carboxyl side of Lys and Arg residues |
| Formulation | Liquid in PBS with EDTA, sterile filtered |
| Appearance | Clear, transparent to slightly colored solution |
| Purity | ≥95% (determined by HPLC — identity confirmed) |
| Activity (1x) | 0.6–1.2 kU/L (photometric; 37 °C, 8 mM L-AAPA, pH 8.0; 1 U = 1 μmol p-nitroaniline/min) |
| Activity (10x) | 6–12 kU/L |
| pH | 7.1–7.6 (Ph. Eur. 2.2.3) |
| Osmolality | 270–320 mOsm/kg (Ph. Eur. 2.2.35) |
| Endotoxin Level (1x) | ≤1 EU/mL (LAL-Test; Ph. Eur. 2.6.14 / USP <85> Method D) |
| Endotoxin Level (10x) | ≤10 EU/mL |
| Mycoplasma Testing | Negative (qPCR Detection Assay; Ph. Eur. 2.6.7) |
| Sterility | Pass (Ph. Eur. 2.6.1 / USP <71>) |
| Storage | 2–8 °C ±5 °C, protected from light |
| Shipping | Ambient temperature (stability data confirmed for ≥6 months) |
| Available Sizes | 100 mL · 500 mL |
| Quality Grades | GMP-grade (EXCiPACT® GMP/GDP) · R&D-grade (ISO 9001) |
| Concentration Formats | 1x ready-to-use · 10x concentrated stock |
| Animal Origin-Free | Yes — raw materials, equipment, and packaging AOF/TSE/BSE certified |
| Antibiotic-Free | Yes |
| Packaging | Non-pyrogenic PET containers, HDPE screw caps; tamper-evident shrink seal (USP Class VI / 21 CFR compliant) |
Validated Performance
CellTrypase has been validated across key biopharma production cell lines for viral vector and vaccine manufacturing, demonstrating efficient trypsinization with high cell viability and robust recovery. The enzyme shows dissociation kinetics, cell replating efficiency, proliferation, and long-term culture maintenance comparable to porcine trypsin and existing recombinant alternatives.
CellTrypase supports direct 1:1 substitution in existing trypsinization protocols. Validated applications include:
- Passaging and expansion of adherent cell lines (e.g., Vero, MDCK, HEK293)
- Cell dissociation in viral vector and vaccine production for cell and gene therapies
- Stem cell processing: iPSCs, hESCs, and organoids
- Primary cell isolation: fibroblasts, keratinocytes, chondrocytes, neuronal cells
- Cell banking and tissue engineering workflows
Inactivation & Handling Protocol
CellTrypase requires no special inhibitor for inactivation. After cell detachment, quench activity by adding fresh medium (serum-free or serum-containing). For complete enzyme removal, centrifuge and resuspend cells in fresh medium.
Quick Protocol (Adherent Cell Dissociation):
- Pre-warm enzyme and medium to 37 °C
- Rinse cells with PBS (Ca²⁺/Mg²⁺-free)
- Add CellTrypase (~1 mL per 25 cm²)
- Incubate at 37 °C (or room temperature) until detachment
- Quench with fresh medium; centrifuge if required
Regulatory Support & Documentation
c-LEcta provides comprehensive documentation to support customer qualification, validation, and regulatory submissions, including: Certificate of Analysis (CoA), Safety Data Sheet (SDS), animal origin-free declarations, TSE/BSE statements, packaging details, change notifications, and quality agreements. A Validation Guide and Application Note for key biopharma cell lines are available upon request. A Drug Master File (DMF) submission is part of the regulatory roadmap for future global filings (EMA, FDA, ANSM).
- CellTrypase Product Information Sheet v2.0 (PDF)
- CellTrypase GMP-Grade 1x – Product Specification (100 mL / 500 mL) v2.0 (PDF)
- CellTrypase R&D-Grade 1x – Product Specification (100 mL / 500 mL) v2.0 (PDF)
- CellTrypase GMP-Grade 10x – Product Specification (100 mL / 500 mL) v2.0 (PDF)
- CellTrypase R&D-Grade 10x – Product Specification (100 mL / 500 mL) v2.0 (PDF)
CellTrypase is a recombinant, animal origin-free trypsin-like enzyme manufactured under GMP conditions (EXCiPACT® GMP/GDP certified). Key advantages include: ≥95% purity verified by HPLC per batch; consistent enzymatic performance across lots; gentler cell dissociation due to lower non-specific reactivity of the fungal enzyme; no requirement for soybean trypsin inhibitor during inactivation; and full AOF/TSE/BSE-free documentation for regulatory submissions. It supports 1:1 direct substitution in existing trypsinization protocols.
Yes. CellTrypase is available in a GMP-grade specifically developed for clinical and commercial biomanufacturing. It is manufactured and distributed under a Quality Management System in compliance with Good Manufacturing and Distribution Practice (GMP/GDP) according to the EXCiPACT® Certification Standard for pharmaceutical excipients — an internationally recognized standard accepted by EMA, FDA, and ANSM. Comprehensive regulatory documentation is available including CoA, SDS, AOF declarations, TSE/BSE statements, and quality agreements. A Drug Master File (DMF) is part of the regulatory roadmap.
CellTrypase has been validated across key biopharma production cell lines used in viral vector and vaccine manufacturing. Validated cell lines include Vero, MDCK, and HEK293 (among others listed in the Application Note available upon request). The enzyme demonstrates efficient dissociation, high cell viability, robust recovery, and comparable long-term proliferation kinetics to the current industry standard. Additional performance data on iPSCs and other biopharma cell lines are available in the Application Note.
Yes. CellTrypase is designed as a direct alternative to TrypLE™ Select and TrypLE™ Express (Thermo Fisher Scientific) as well as recombinant porcine trypsin solutions. The GMP-grade matches TrypLE™ Select performance, while the R&D-grade matches TrypLE™ Express. CellTrypase is compatible with existing trypsinization protocols, enabling 1:1 substitution without process revalidation. A regulatory-aligned Guidance Document for qualification and switching scenarios is available upon request from c-LEcta.
CellTrypase should be stored at 2–8°C (±5°C), protected from light, for long-term stability. Stability data confirm consistent enzymatic activity under refrigerated, frozen, and ambient storage conditions for at least 6 months. The product's superior stability as a liquid formulation supports ambient temperature shipping — no cold-chain is required. Short-term room temperature storage (at least 1 month) does not result in measurable activity loss, although the enzyme is light-sensitive and should be protected from light at all times.
CellTrypase is available in GMP-grade and R&D-grade formulations in both 1x ready-to-use and 10x concentrated stock formats, with 100 mL and 500 mL bottle sizes. A free 100 mL sample is available for process development, testing, and qualification — contact the c-LEcta expert team to request yours. For volume pricing, custom packaging inquiries, or bulk supply requirements, please reach out to our specialist team via the product contact form. GMP-grade orders include dedicated regulatory support for pharmaceutical product approvals.
