CellTrypase – Recombinant Trypsin-Like Enzyme for Cell Dissociation

Scientific background

Trypsin-like serine proteases cleave peptide bonds on the carboxyl side of lysine and arginine residues — the same chemistry exploited during cell dissociation. Conventional porcine trypsin carries the biosafety considerations associated with animal-derived materials, including lot-to-lot variability and the theoretical risk of adventitious agents. Recombinant, non-mammalian alternatives remove the animal origin entirely and are produced under controlled fermentation conditions.

CellTrypase is a recombinant trypsin-like enzyme derived from Fusarium oxysporum and produced by microbial fermentation using Bacillus sp. as the expression host. Its fungal origin confers lower non-specific reactivity than mammalian trypsin, and the calculated molecular weight of the single-chain enzyme is approximately 22 kDa¹.

Product description

CellTrypase is manufactured in-house at the Kerry Biotechnology Centre in Leipzig, Germany, by c-LEcta GmbH — now operating as Kerry (formerly c-LEcta) — under a certified quality management system. It is available in two grades — a GMP grade produced under a QMS compliant with the EXCiPACT® certification standard for pharmaceutical excipients, and an R&D grade manufactured under ISO 9001 — and in two concentration formats, 1× ready-to-use and 10× concentrated stock, supplied in 100 mL and 500 mL bottles. Both grades are made by the same process and are released against the same specification.

The formulation is a sterile-filtered liquid in PBS with 1.1 mM EDTA. Supplying the enzyme as a liquid removes drying and reconstitution steps, which preserves enzyme integrity and gives a clear, ready-to-use solution.

Measured dissociation performance

Kerry tested the 1× working solution on four adherent production cell lines over five consecutive passages (n = 3 technical replicates), measuring dissociation time, viability, cell yield and population doubling time against the current industry-standard trypsin-like enzyme as the control. The comparator is not named in the source documents; it is described only as the "current industry standard trypsin-like enzyme".

Mean viability and yield are averages across five passages (P0–P4); yield is expressed relative to the industry-standard control. MDCK is a tightly adherent line and takes over 20 minutes with either enzyme. Source: Kerry scientific poster "Advancing cell dissociation in bioprocessing with CellTrypase" (Feb 2026) and CellTrypase Product Information Sheet v4.0 (Aug 2026).

Kerry also reports data in human induced pluripotent stem cells (iPSCs), the most sensitive system tested. In 2D colonies, 1× CellTrypase reached single-cell suspension in 5 minutes versus 7 minutes for the control, with 94% viability for both enzymes, 99% relative yield, residual aggregates below 5%, and pluripotency marker expression (Oct3/4, SSEA4, TRA-1-60, CD30, Nanog, Sox2, SSEA3) above the 75% acceptance threshold by flow cytometry. In 3D pluripotent aggregates from bioreactor culture, both enzymes required about 10 minutes; CellTrypase gave 87% viability (control 85%), 104% relative yield and fewer residual aggregates (12% versus 16%).

1× CellTrypase versus the industry-standard control; mean ± SD of n = 3 technical replicates. Source: Kerry poster, Feb 2026. Data on differentiated 3D iPSC aggregates and the full five-passage figure set for each cell line are in the CellTrypase Application Note v1.1, which Kerry supplies on request — request it here.

CellTrypase requires no trypsin inhibitor for inactivation — quenching by dilution in fresh medium is sufficient. Every lot is released at ≥95% purity by size-exclusion HPLC; the chromatogram of the 10× GMP stock shows a single, well-defined peak, whereas the industry-standard comparator shows a broader peak base with small additional signals (see Validation & QC below).

Intended applications

Kerry specifies CellTrypase for gentle, efficient dissociation of adherent and aggregating cells in biopharmaceutical manufacturing and research, including:

• Passaging and expansion of adherent production lines such as CHO-K1, HEK 293, Vero and MDCK
• Cell dissociation in viral vector and viral vaccine production for cell and gene therapies
• Passaging of human iPSCs in 2D and 3D culture, with pluripotency marker expression preserved
• Sensitive cell types such as hESCs, primary cell isolation, organoid and 3D culture models, cell banking and differentiation workflows

Released comparative data cover CHO-K1, HEK 293, MDCK, Vero and human iPSCs (2D, 3D and differentiated 3D aggregates). Performance on other cell types — hESC, primary cells, organoids, MSCs and others — is stated by the manufacturer as an intended use but is not backed by published figures. As with any change of raw material, qualify the enzyme on your own cells before adopting it in a routine or regulated workflow. A free 100 mL sample is available for exactly that purpose.

Inactivation and handling

CellTrypase requires no special inhibitor. After the cells detach, quench the reaction by adding fresh medium, serum-free or serum-containing. For complete enzyme removal, centrifuge and resuspend the cells in fresh medium. Residual enzyme can be quantified with commercially available ELISA kits for recombinant fungal trypsin-like enzymes; Kerry has experimentally confirmed compatibility with these assay formats.

Quick method for adherent cells:

• Pre-warm the enzyme and medium to room temperature or 37 °C — warmer temperatures accelerate detachment
• Remove spent medium and rinse the monolayer with Ca²⁺/Mg²⁺-free PBS
• Add approximately 1 mL of the 1× solution per 25 cm²
• Incubate until the cells detach — check the monolayer rather than relying on a fixed time; tap the vessel or pipette gently if needed
• Quench with fresh medium; centrifuge if the enzyme must be removed

The 10× concentrate may be used undiluted for fast detachment of strongly adherent cells or diluted to 1× in PBS containing 1 mM EDTA under sterile conditions. The enzyme is most active between pH 7.0 and 10.0 and between 20 and 37 °C; activity falls sharply above 60 °C and is minimal below pH 6.

Storage and shipping

Store at 2–8 °C, protected from light. Kerry's Product Information Sheet v4.0 (August 2026) confirms stability for at least 21 months from the date of manufacture at 2–8 °C for both the GMP and R&D grades, based on real-time stability data; long-term stability is still under investigation and the shelf life is extended as data accumulate. Temporary exposure to room temperature or above during shipment does not affect enzyme activity, so the product ships at ambient temperature and no cold chain is required. Avoid extended storage at or near 37 °C, which causes a gradual loss of activity through autoproteolysis.

Regulatory support and documentation

Kerry provides documentation to support customer qualification, validation and regulatory submissions, including Certificate of Analysis, Safety Data Sheet, animal origin-free declarations, TSE/BSE statements, packaging details, change notifications and quality agreements. Both grades are manufactured without antibiotics, animal-derived raw materials or TSE/BSE-risk materials; the production host is a GMO of safety level S1 and the final product contains no GMO. The Product Information Sheet v4.0 and the four v4.0 product specifications are available below. The CellTrypase Validation Guide v1.2 (release test methods, stability and regulatory statements) and the Application Note v1.1 (full performance data) are supplied on request through the technical documents page.

¹ Calculated using Clone Manager 12 Professional Edition (Sci Ed Software); reflects calculation according to Lehninger. Unit definition: one unit (U) forms 1 µmol p-nitroaniline per minute at 37 °C from 8 mM L-AAPA at pH 8.0.

For Research Use Only. GMP-grade material is intended as a processing aid in biopharmaceutical manufacturing and is not for direct administration to humans or animals. Manufactured and distributed by c-LEcta GmbH, Kerry Biotechnology Centre, Leipzig, Germany — Kerry (formerly c-LEcta). CellTrypase is a trademark of Kerry Group. Free 100 mL samples are available for qualification and process-development testing.