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सी.एस.आई.आर - केन्द्रीय औषधि अनुसंधान संस्थान, लखनऊ
CSIR-Central Drug Research Institute, Lucknow

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Bio-Layer Interferometry (BLI)

Label free technology for the detection of bio-molecular interaction

Octet R8 (BLI - Bio-Layer Interferometry) simultaneously collect data from up to eight “dip-and-read” sensors. This instrument uses 96-well plates for sample handling. The molecule of interest ("ligand") is immobilized on the surface of a sensor, creating the biolayer. When this sensor is dipped into a well containing a solution of the ligand’s binding partner ("analyte"), the analyte molecules bind to the immobilized ligand molecules, changing the biolayer’s thickness. To monitor the biolayer thickness changes, Octet analyzes the interference of the light beam reflected from the tip of the sensor. In this way, Octet can track the association and dissociation of the analyte from the immobilized ligand in real time. It can use crude sample solutions, such as cell culture supernatants, diluted serum, etc. The BLI signal is sensitive to the binding of a large range of analytes, from small molecules (MW > 150 Da) to virus particles. Most sensors can be regenerated and used multiple times.

Sample requirements
  1. A minimum of 400 μL of both the ligand and analyte solutions is required to set up the plate with the necessary controls (a binding-check experiment).
  2. All buffers (Kinetic buffer, regeneration buffer and neutralization buffer) should be prepared in deionized water and degassed for 15 minutes.
  3. Sample and buffer should be filtered through 0.22µM filter unit.
  4. Please ensure that samples are not aggregated.
  5. The ligand concentration required for loading sensors should be standardized varying from 5µg/ml to 50µg/ml or higher.
  6. The analyte solution (solution of molecules interacting with the immobilized ligands) should be available at a concentration of at least 10x its dissociation constant value; if the dissociation constant is unknown, we recommend a concentration of 1 μM or higher.
  7. All the consumables including biosensors should be brought by the user and only the access to the instrument will be provided from CSIR-CDRI end. Technical person involved will be operating and collecting the data.

Contact:

Dr. Kaushik Bhattacharjee

Coordinator, Biological Screening

+91-522-2772486

bio-screen[AT]cdri.res.in

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CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow-226031

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