Activity assay for bond forming enzymes
AssigneeHoffmann-La Roche Inc.
InventorsMara Boenitz-Dulat, Erhard Kopetzki, Peter Kratzsch, Martin Schatte
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Abstract
Herein is reported a method for the detection of a sortase in a sample, comprising the following steps: a) incubating the sample with a first substrate comprising an immobilization tag and a second substrate comprising a detectable label, whereby in the presence of a sortase in the sample a conjugate comprising the immobilization tag and the detectable label is formed, b) immobilizing the conjugate of step a) via/using the immobilization tag to a solid phase, c) detecting the immobilized conjugate via/using the detectable label and thereby detecting the sortase in the sample.
Claims32
A method for the detection of a bond forming enzyme in a sample, comprising the following steps:
- a) incubating the sample in a solution with a first substrate comprising an immobilization tag and a second substrate comprising a detectable label, whereby in the presence of a bond forming enzyme in the sample a conjugate comprising the immobilization tag and the detectable label is formed,
- b) centrifuging the solution from step a) and collecting a supernatant,
- c) immobilizing the conjugate in the supernatant of step b) using the immobilization tag to a solid phase, and
- d) detecting the immobilized conjugate using the detectable label, thereby detecting the bond forming enzyme in the sample.
Dependencies are inferred from the claim text.