膜杰作
Star Staining
描述（Description）
The Human CD16a (158V) (Luc) Jurkat Reporter Cell was engineered to not only express the NFAT response element driving luciferase expressing systems, but also express full length human CD16a receptor mutated to a Valine (V) at amino acid 158 exhibiting a higher affinity for IgG1 and IgG3 isotypes compared to CD16a-158F, which can use to evaluate ADCC activity of antibodies in the presence of corresponding target cells. When co-cultured with a target cell and relevant antibody, the antibody simultaneously binds the target cell antigen and CD16a (158V) receptor on the surface of Human CD16a (158V) (Luc) Jurkat Reporter Cell, resulting in receptor clustering, intracellular signaling and NFAT-mediated luminescence.
应用说明（Application）
Determination of ADCC activity induced by antibodies
生长特性（Growth Properties）
Suspension
筛选标记（Selection Marker）
Puromycin (5 μg/mL) + Hygromycin (20 μg/mL)
培养基（Culture Medium）
RPMI-1640 + 10% FBS
冻存液（Freeze Medium）
Serum-free cell cryopreservation medium
装量（Quantity）
1 vial contains at least 5×10^6 cells in 1 mL serum-free cryopreservation medium
存储（Storage）
Frozen in liquid nitrogen.
支原体检测（Mycoplasma Testing）
Negative
无菌检测（Sterility Testing）
Negative
使用说明（Instructions for Use）
See data sheet for detailed culturing and assay protocol.
Receptor Assay
Expression analysis of human CD16a (158V) on Human CD16a (158V) (Luc) Jurkat Reporter Cell by FACS.
Human CD16a (158V) (Luc) Jurkat Reporter Cell or negative control cell were stained with PE-labeled anti-human CD16a antibody.
Protocol
Application
ADCC response to anti-human CD20 antibody (RLU).
Anti-human CD20 antibody-induced ADCC activity was evaluated using Human CD16a (158V) (Luc) Jurkat Reporter Cell in the presence of Raji cells that express CD20 endogenously. The EC50 of anti-human CD20 antibody was approximately 0.0028 μg/mL.
Protocol
ADCC response to anti-human CD20 antibody (FOLD).
Anti-human CD20 antibody-induced ADCC activity was evaluated using Human CD16a (158V) (Luc) Jurkat Reporter Cell in the presence of Raji cells that express CD20 endogenously. The max induction fold was approximately 854.
Protocol
Passage Stability
Passage stability analysis by Signaling Bioassay.
The continuously growing Human CD16a (158V) (Luc) Jurkat Reporter Cell was stimulated with serial dilutions of anti-human CD20 antibody in the presence of Raji cells that express CD20 endogenously. Anti-human CD20 antibody stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 14-26.
Protocol
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背景（Background）
Antibody-dependent cell-mediated cytotoxicity (ADCC) is a potent cytotoxic mechanism that is mainly mediated in humans by natural killer (NK) cells. ADCC mediates the clinical benefit of several widely used cytolytic monoclonal antibodies (mAbs), and increasing its efficacy would improve cancer immunotherapy. CD16a is a receptor for the Fc portion of IgGs and is responsible to trigger NK cell-mediated ADCC. Previous study indicated that t, CD16a 158V shows a better ADCC in vitro. Acrobiosystems offers a series of NFAT (Luc) Jurkat Reporter Cell specifically designed to assess the potency of specific immunoglobulin for (antibody-dependent cellular cytotoxicity) and (antibody-dependent cell-mediated phagocytosis). These cells derive from the human T lymphocyte Jurkat cell line and stably express CD16 or CD32, two Fc-gamma receptors (FcγR) for the constant region of immunoglobulin G (IgG).
License Disclosure
This cell line is provided for research use only. This license does not permit you to share, distribute, sell, sublicense, or otherwise make this cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. The license does not permit modification of this cell line in any way. Inappropriate use or distribution of this cell line will result in revocation of the license. Modifications of this cell line, transfer to another facility, or commercial use of the cell lines may require a separate license and additional fees. AcroBiosystems does not warrant the suitability of this cell line for any particular use, and does not accept any liability in connection with the handling or use of this cell line.
