Method for isolating cell nuclei having enhanced antigenicity from fixed cells or FFPE tissue section, and antigen activator and kit therefor
US12153054B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Feb 28, 2019 |
| Grant date | Nov 26, 2024 |
| Priority date | — |
| Expiry date | Dec 25, 2041 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2800/52
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
Provided are a pretreatment method for detecting the number of cells containing Ki-67 protein-positive nuclei using Ki-67 antibody; a method for the detection; a kit to be used in the detection method; and determination of a therapy regimen using the aforesaid method. Attempts were made to activate Ki-67 antigen with the use of an enzyme having been considered as inappropriate for the activation thereof. By pretreating a sample with an enzyme not recognizing the epitope of MIB-1 (a rare cutter enzyme), the antigen activation of Ki-67 antigen was enhanced, while enhancing the antigenicity of other antigens including a cytokeratin too. As a result, a method for more objectively and more universally quantifying Ki-67-positive cells at higher reproducibility, said method comprising isolating cell nuclei from an FFPE section while enhancing the antigenicity and performing a reaction between Ki-67 protein, i.e., the target, existing in the cell nuclei with a fluorescently labeled antibody, has been completed.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.