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US 9,663,814 B2Granted
Proximity assay for in situ detection of targets
AssigneeVentana Medical Systems, Inc.
InventorsMichael Farrell, Rui Hong, Zeyu Jiang
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AssigneeVentana Medical Systems, Inc.
InventorsMichael Farrell, Rui Hong, Zeyu Jiang
Estimating the term…
A proximity detection method is described that utilizes enzymatic biotinylation to detect targets in a sample, particularly formalin fixed paraffin embedded samples using automated staining platforms. One disclosed embodiment comprises contacting the sample with a first conjugate comprising a biotin ligase and a first specific binding moiety that binds proximally to the first target; contacting the sample with a second conjugate comprising a biotin ligase substrate and a second specific binding moiety that binds proximally to the second target; subjecting the sample to conditions that allow biotinylation of the biotin ligase substrate by the biotin ligase when the first target and the second target have a proximal arrangement; and detecting biotinylation of the biotin ligase substrate. The conditions that allow biotinylation of the substrate include addition of biotin and ATP. The method also may comprise contacting the sample with a streptavidin-enzyme conjugate. Signal amplification also can be used.
A method for analyzing a sample to determine whether a first target is proximal to a second target, the method comprising:
An automated method, comprising using an automated staining apparatus to perform one or more steps associated with a method of testing a formalin fixed paraffin embedded tissue sample for the presence of a first target and a second target in close proximity, the method comprising:
Dependencies are inferred from the claim text.