Raji (HT-HiBiT) TCK Bioassay Technical Manual
Instructions for Use of Product(s)
JA1211, JA1215, JA2441, JA2445, JA1251, JA1255, JA1261, JA1265, JA1271, JA1275
Literature # TM733
Inducing tumor cell lysis by immune effector cells is a primary mechanism of action (MOA) for many immuno-oncology drugs. Monoclonal antibodies against tumor-associated antigens (TAA) can induce antibody-dependent cellular cytotoxicity (ADCC) by natural killer (NK) cells. Next-generation biologics, such as bispecific antibodies, can redirect T cells to lyse tumor cells. CAR-T cells and other cellular therapies are living drugs that directly lyse tumor cells as a component of their MOA. HiBiT Target Cell Killing (TCK) Bioassays provide a flexible, reproducible, label-free luminescent system for measuring the potency of a variety of cellular and immuno-oncology drug products where target cell killing is a MOA.
Raji (HT-HiBiT) TCK Bioassays measure 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.
This Technical Manual is relevant for Cat.# JA1211, JA1215, JA1251, JA1255, JA1261, JA1265, JA1271 and JA1275; JA1641, JA1645, JA1651, JA1655, JA1661, JA1665, JA2441 and JA2445 (Korea).
Summary of Changes
The following changes were made to the 1/25 revision of this document:
1. Six new Raji (HT-HiBiT) Bioassay Kits including knockout (KO) cells were added to Section 2, as well as to the protocol sections.
2. Bio-Glo-NB™ Lytic Luciferase Assay, Cat.# JB1101, was added.
3. Figures 1 and 2 were updated. Figure 6 was added.
4. Limited Use Label Licenses were added and updated, one patent statement updated and a patent statement was removed.
5. Trademarks were updated.
Revised 1/25.