公开(公告)号：US20190338373A1

公开(公告)日：2019-11-07

申请号：US16401906

申请日：2019-05-02

Applicant: INDIAN INSTITUTE OF SCIENCE EDUCATION AND RESEARCH

Inventor： Sandanaraj Selvaraj BRITTO ,  Punita BATHLA

IPC: C12Q1/6897 ,  C12Q1/37 ,  C12Q1/02

Abstract: Methods and materials for specific imaging of active enzyme in a live or intact cell are disclosed. The enzyme of interest tagged to reporter protein (donor) is exogenously expressed in a cell. The conversion of proenzyme to active enzyme (containing reporter protein) is achieved upon applying an appropriate stimulus to the target cells. The activated enzyme is labelled with an activity-based probe carrying a fluorophore (acceptor). The covalent labelling of active enzyme by the activity-based probe creates a FRET pair which provides the opportunity to exquisitely image the function of an “active enzyme”. This method is used for specific imaging of the function of active caspase-3,-7,-8,-9 and cathepsin-B and also for profiling of inhibitors of caspases and cathepsin B.

公开(公告)号：US20220298541A1

公开(公告)日：2022-09-22

申请号：US17698397

申请日：2022-03-18

Applicant: Indian Institute of Science Education and Research

Inventor： Britto S. SANDANARAJ ,  Punita BATHLA ,  Abhijit DE ,  Aaiyas MUJAWAR

IPC: C12Q1/66 ,  G01N33/58 ,  G01N33/50

Abstract: The present disclosure relates to an assay for determining drug-target engagement in real-time. Specifically, the present disclosure provides a method for determining drug candidate-target interaction in real-time in living systems using Activity-based Reporter Gene Technology-Bioluminescence Resonance Energy Transfer (AbRGT-BRET) based assay. The said assay is useful for High Throughput Screening of compounds to a specific target.