H929 (HiBiT) TCK Bioassay Technical Manual
Instructions for Use of Product(s)
JA1511, JA1515, JA2471, JA2475
Literature # TM747
The H929 (HiBiT) TCK Bioassay measures the death of target cells engineered to express a HiBiT fusion protein. HiBiT is an 11 amino acid peptide that binds with high affinity to LgBiT to form NanoBiT® luciferase. When dead or dying cells release the HiBiT fusion protein into the extracellular medium, it binds to the cell impermeable LgBiT, which is provided with the furimazine (Fz) substrate in a nonlytic, homogeneous detection reagent. In coculture experiments, the luminescent signal will be proportional to target cell death alone, with no signal contribution arising from the death of effector cells.
H929 (HiBiT) TCK Bioassay is provided in a thaw-and-use format as cryopreserved cells that can be thawed, plated and used in an assay without the need for cell culture and propagation.
When H929 (HT-HiBiT) target cells are cocultured with appropriate effector cells (e.g., NK cells or cytotoxic T cells) they can be used to specifically monitor the activity of biologics directed against receptors expressed on the H929 Cells (HT-HiBiT), such as CD38 or BCMA. When H929 Cells (HT-HiBiT) are used in assays with biologics against TAAs they do not express, the target cells are not lysed and no increase in luminescent signal is observed.
This Technical Manual is relevant for Cat.# JA1511 and JA1515; JA2471 and JA2475 (Korea).
The following changes were made to the 3/25 version of this document:
1. In Section 2, quantities of RPMI 1640 and FBS supplied in the kits have changed.
2. In Section 3, text about the product label was revised.
3. The amount of assay buffer to use in Section 5.E, Step 6 was increased.
4. Some subheadings were updated to include the day on which that protocol section is performed.
5. In Section 4, RPMI 1640 medium and FBS were added to the Materials to Be Supplied list.
6. Two patent statements were removed.
Revised 3/25.