Product Details
- Genetically modified cell lines best reflect MOA (Mechanism of Action)
- Higher activity and larger assay window for robust and reproducible cell-based bioassay
- Comprehensive application data to support assay development and validation
- Full tracible record, stringent quality control and validated cell passage stability
- Parental cell line legally obtained from internationally recognized cell resource bank and commercially licensed
- Global commercial license assistance whenever regulatory filing is required
Description
The Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) was engineered to not only express the NFAT signaling response element and the receptor full length human CD16a (Uniprot: P08637) mutated to a Valine (V) at amino acid 158 exhibiting a higher affinity for IgG1 and IgG3 isotypes compared to CD16a-158F, but also targeted knockout of the endogenous TCR chains resulting in loss of functional TCR/CD3 complex on the cell surface. When co-cultured with a target cell and the corresponding anti-TAA × CD3 bispecific antibody (BsAb), the anti-TAA × CD3 BsAb simultaneously binds both the target-associated antigen (TAA) on the target cell and the CD16a (158V) receptor on the Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative), resulting in receptor clustering, intracellular signaling and NFAT-mediated luminescence.
Application
• Determination of ADCC activity induced by anti-TAA × CD3 BsAbs

Growth Properties
Suspension
Selection Marker
Puromycin (5 μg/mL) + Hygromycin (20 μg/mL) + Zeocin (20 μg/mL)
Complete Growth Medium
RPMI-1640 + 10% FBS
Freeze Medium
Serum-free cell cryopreservation medium
Quantity
1 vial contains at least 5×10^6 cells in 1 mL serum-free cryopreservation medium
Storage
Frozen in liquid nitrogen.
Mycoplasma Testing
Negative
Sterility Testing
Negative
Instructions for Use
See data sheet for detailed culturing and assay protocol.
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Performance Data
Receptor Assay

Expression analysis of human CD16a (158V) on Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) by FACS.
Cell surface staining was performed on Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) or negative control cell using PE-labeled anti-human CD16a antibody.Protocol
Expression analysis of human TCR and human CD3 on Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) by FACS.
Cell surface staining was performed on Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) using PE-labeled anti-human TCR antibody and APC-labeled anti-human CD3 antibody. The parental reporter cell (CD3 Positive) was used as the control. This result confirmed the loss of TCR/CD3 complex on the cell surface of Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative).Protocol
Signaling Bioassay

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 (CD3 Negative) in the presence of Raji cells that express CD20 endogenously.The parental reporter cell lacking human CD16a (158V) expression was used as the control. The EC50 of anti-human CD20 antibody on the Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) was approximately 0.0038 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 (CD3 Negative) in the presence of Raji cells that express CD20 endogenously. The parental reporter cell lacking human CD16a (158V) expression was used as the control. The max induction fold on Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) was approximately 718.04.Protocol
Response to anti-human CD3 antibody (RLU).
Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) was incubated with serial dilutions of anti-human CD3 antibody (Cat. No. CDE-M120a). The parental reporter cell (CD3 Positive) was used as the control. Compared to the parental reporter cell (CD3 Positive), the Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) exhibited no activation response to anti-human CD3 antibody stimulation.Protocol
Response to anti-human CD3 antibody (FOLD).
Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) was incubated with serial dilutions of anti-human CD3 antibody (Cat. No. CDE-M120a). The parental reporter cell (CD3 positive) was used as the control. Compared to the parental reporter cell (CD3 Positive), the Human CD16a (158V) (Luc) Jurkat Reporter Cell (CD3 Negative) exhibited no activation response to anti-human CD3 antibody stimulation.Protocol
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Important Statement
Limited Use&License Disclosure
BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE FOLLOWING TERMS OF LIMITED USE OF THIS CELL LINE PRODUCT.
- If the researcher is not willing to accept the terms of limited use of this cell line product, and the product is unused, ACRO will accept return of the unused product.
- Researchers may use this product for research use only, no commercial use is allowed. "Commercial use" means any and all uses of this product and derivatives by a party for profit or other consideration and may include but is not limited to use in: (1) product manufacture; and (2) to provide a service, information or data; and/or resale of the product or its derivatives, whether or not such product or derivatives are resold for use in research.
- This cell line is neither intended for any animal or human therapeutic purposes nor for any direct human in vivo use. You have no right to share, modify, transfer, distribute, sell, sublicense, or otherwise make the cell line available for use to other researchers, laboratories, research institutions, hospitals, universities, or service organizations.
- ACROBIOSYSTEMS MAKES NO WARRANTIES OR REPRESENTATIONS OF ANY KIND, EITHER EXPRESSED OR IMPLIED, WITH RESPECT TO THE SUITABILITY OF THE CELL LINE FOR ANY PARTICULAR USE.
- ACROBIOSYSTEMS ACCEPTS NO LIABILITY IN CONNECTION WITH THE HANDLING OR USE OF THE CELL LINE.
- Modifications of the cell line, transfer to a third party, or commercial use of the cell line may require a separate license and additional fees. Please contact order@acrobiosystems.com for further details.
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