CellTrypase is a recombinant, animal-origin-free trypsin-like enzyme for dissociating adherent cells — a defined, 1:1 drop-in for porcine trypsin and TrypLE™. This application note summarizes how it compares on dissociation performance, enzyme purity, and positioning, so you can decide where it fits in your workflow.
Why move beyond porcine trypsin
Conventional porcine trypsin carries animal-derived biosafety risk (TSE/BSE and adventitious agents), batch-to-batch variability, and requires a trypsin inhibitor to stop the reaction. Animal-free recombinant trypsin-like enzymes — including TrypLE™ and CellTrypase — remove those liabilities while supporting a 1:1 swap into existing trypsinization protocols.
What CellTrypase is
CellTrypase is a recombinant Fusarium oxysporum trypsin-like serine protease produced by microbial fermentation. It cleaves on the carboxyl side of lysine and arginine residues, is purified to ≥95% by HPLC per lot, and requires no inhibitor — activity is quenched by dilution in medium. It is supplied as a ready-to-use 1× solution or a 10× concentrated stock, in 100 mL and 500 mL sizes, and is available in two grades from a single source: an R&D grade (in-house, ISO 9001-compliant manufacturing) and a GMP grade (in-house GMP production under a QMS compliant with the EXCiPACT® certification standard). It ships at ambient temperature and is stable for at least 15 months at 2–8 °C. For research use only (RUO); the GMP grade is a biomanufacturing processing aid, not for direct human or animal administration. Source: CellTrypase Product Information Sheet v2.0 and c-LEcta product literature.
Head-to-head dissociation performance
In c-LEcta’s validation, CellTrypase (1× working solution) matched the current industry-standard trypsin-like enzyme on dissociation, with high viability and equal-or-higher cell yield:
| Cell line | Release time | Mean viability | Yield vs standard |
|---|---|---|---|
| CHO-K1 | 2:25 | 98% | 106% |
| HEK 293 | 2:27 | 95% | 100% |
| MDCK | 23:32 | 99% | 102% |
| Vero | 4:07 | 99% | 103% |
Yield is expressed as a percentage of the current industry-standard trypsin-like enzyme (control). Source: CellTrypase PIS v2.0 (c-LEcta).
Enzyme purity and consistency
In c-LEcta’s HPLC analysis, CellTrypase resolves as a single, sharp peak with minimal by-products — a defined, single-enzyme preparation that supports consistent, low-off-target dissociation batch to batch. (TrypLE™ Select is likewise marketed as a high-purity, single-enzyme reagent.)
How it compares to TrypLE™
For most bench dissociation, CellTrypase and TrypLE™ are close substitutes. Both are animal-free, recombinant fungal trypsin-like enzymes; both are defined, require no inhibitor (inactivated by dilution), are stable at ambient/room temperature, and are available in 1× and 10× formats.
| Attribute | CellTrypase | TrypLE™ |
|---|---|---|
| Origin | Recombinant F. oxysporum (fermentation) | Recombinant fungal protease (fermentation) |
| Animal-free | Yes | Yes |
| Inhibitor needed | No (dilute to quench) | No (dilute to quench) |
| Defined / purity | ≥95% HPLC, single-enzyme | High-purity, single-enzyme |
| Formats | 1×, 10× · 100/500 mL | 1×, 10× · multiple sizes |
| Grades | R&D (ISO 9001) + GMP (QMS per EXCiPACT® standard) | Express, Select, CTS (DMF) |
| Free trial | Yes — free 100 mL | — |
The practical differences are grade fit, price, and the option to trial CellTrypase free. CellTrypase offers R&D and GMP grades from one supplier — the GMP grade produced under a QMS compliant with the EXCiPACT® certification standard; TrypLE™ spans Express (general purpose), Select (research/bioproduction), and CTS (cell-therapy, with Drug Master File support). CellTrypase R&D grade is competitively priced, and a free 100 mL sample lets you validate it on your own cells before switching.
When to use which
- Coming from porcine trypsin: CellTrypase is a clear upgrade — animal-free, defined, no inhibitor, and gentler.
- Currently using TrypLE™: CellTrypase is a comparable alternative; the free 100 mL lets you confirm performance on your cells with no protocol change.
- Sensitive cells (iPSC/hESC, primary cells, organoids): a defined, low-off-target enzyme helps preserve viability and surface markers.
Ready to run it on your cells? Follow the step-by-step CellTrypase Cell Dissociation Protocol — pre-warming, working amounts, 10× concentrate use, and troubleshooting.
Specifications at a glance
| Enzyme class | Serine protease (trypsin-like); cleaves after Lys & Arg |
| Source / expression | Recombinant Fusarium oxysporum, microbial fermentation |
| Molecular weight | ~22 kDa |
| Purity | ≥95% (HPLC) |
| Activity | 0.6–1.2 kU/L (1×); 6–12 kU/L (10×) |
| Endotoxin | ≤1 EU/mL (1×) |
| Mycoplasma / sterility | Negative / Pass |
| pH / osmolality | 7.1–7.6 · 270–320 mOsm/kg |
| Storage / shipping | 2–8 °C; ships ambient; ≥15-month stability |
| Formats | 1× ready-to-use, 10× stock · 100/500 mL |
| Grades | R&D (ISO 9001), GMP (QMS compliant with EXCiPACT® standard) |
Source: CellTrypase Product Information Sheet v2.0 and c-LEcta product literature. For research use only.
Request a free 100 mL sample
BioHippo offers a free 100 mL CellTrypase sample for qualification and process-development testing — one per lab, while supplies last. Request your free sample or view full specifications on the CellTrypase product page.
