CD40 Bioassay Technical Manual
Instructions for Use of Product(s)
JA2151, JA2155, JA3151, JA3155, JA4151, JA4155, JA2251, JA2255
Literature # TM568
The CD40 Bioassay (Cat.# JA2151, JA2155), is a bioluminescent cell-based assay that overcomes the limitations of existing assays and can be used to measure the potency and stability of ligands or agonist antibodies and other biologics that can bind and activate CD40. The assay consists of a genetically engineered cell line that expresses human CD40 and a luciferase reporter driven by a response element that can respond to CD40 ligand/agonist antibody stimulation. The CD40 Effector Cells are provided in 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. The CD40 Effector Cells are also provided in Cell Propagation Model (CPM) format (Cat.# J2132), as cryopreserved cells that can be thawed, propagated and banked for long-term use.
Based on the antibody properties to be tested, the CD40 Bioassay can be conducted in either single-cell systems or with FcγRIIb CHO-K1 Cells (Cat.# JA2251, JA2255) to determine the agonist antibodies that can activate CD40 in a FcγR-dependent manner. Induction of the CD40 Effector Cells with CD40 ligand or an agonist CD40 antibody results in response element-mediated luminescence. The bioluminescent signal is quantified using the Bio-Glo™ Luciferase Assay System and a standard luminometer such as the GloMax® Discover System.
This Technical Manual is relevant for Cat.# JA2151, JA2155, JA2251 and JA2255; JA3151 and JA3155 (Korea); JA4151 and JA4155 (Taiwan).
Summary of Changes:
The following changes were made to the 10/24 revision of this document:
1. Corrected the x axis of Figure 2, Panel B, changing μg/ml to g/ml.
2. Updated the cover image and fonts.
3. Made miscellaneous text edits, including updating Section 2 storage temperatures per our latest style, and without changing temperatures and updating one patent statement and Related Products, Section 8.C.
Revised 10/24.