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Case study · Biopharma & ADC
Results at a glance
>99%
purity after the first purification step
10×
higher unit activity than a comparison commercial enzyme
80%
less enzyme than a CRO expected to need for its conjugations
The challenge
Conjugation is only as good as the enzyme.
Sortase-mediated conjugation depends on enzyme quality. Lower purity and lower activity mean more enzyme per reaction, more variability between lots, and more enzyme-derived impurities to clear from the final conjugate.
What we did
High purity in one step, tested head to head.
We manufacture Sortase A heptamutant with CLīM affinity purification, reaching about 99% purity in the first purification step. We then compared its in vitro activity with Sortase A heptamutant from another commercial supplier at 100, 50 and 25 ng per reaction.
About 99% purity by SDS-PAGE after one step
Activity tested at 100, 50 and 25 ng per reaction
Head to head against a commercial Sortase A
Available from milligrams to recurring bulk supply
Results
Higher activity at every dose.
About 10× the activity
TriAltus Sortase A gave roughly ten times the activity signal of the comparison enzyme across the doses tested.
A quarter of the enzyme, more signal
At 25 ng per reaction, TriAltus Sortase A outperformed the comparison enzyme used at 100 ng.
>99% first-step purity
A single clean band by SDS-PAGE after the first purification step.
Director of Bioconjugation, CRO
Why it matters
What this means for your program.
1
Less enzyme per conjugation
Higher unit activity means less enzyme per reaction, which lowers cost of goods downstream.
2
Cleaner conjugates
High starting purity leaves fewer enzyme-derived impurities to clear.
3
Supply you can build on
Lot traceability and change control support your program from development to supply.
Activity comparison: in vitro fluorescence activity assay of Sortase A heptamutant from TriAltus and from another commercial supplier at 100, 50 and 25 ng per reaction. Purity by SDS-PAGE. Customer name withheld.