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Human CD64 (Luc) Jurkat Reporter Cell Development Service

For research use only.

  1. Genetically modified cell lines best reflect MOA (Mechanism of Action)
  2. Higher activity and larger assay window for robust and reproducible cell-based bioassay
  3. Comprehensive application data to support assay development and validation
  4. Full tracible record, stringent quality control and validated cell passage stability
  5. Parental cell line legally obtained from internationally recognized cell resource bank and commercially licensed
  6. Global commercial license assistance whenever regulatory filing is required

描述(Description)

The Human CD64 (Luc) Jurkat Reporter Cell was engineered to not only express the NFAT response element driving luciferase expressing systems, but also express the receptor human CD64 (Gene ID: 2209), which can use to evaluate ADCP 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 CD64 receptor on the surface of Human CD64 (Luc) Jurkat Reporter Cell, resulting in receptor clustering, intracellular signaling and NFAT-mediated luminescence.

应用说明(Application)

• Determination of ADCP activity induced by antibodies

Fc gamma RI / CD64 Assay Principles

生长特性(Growth Properties)

Suspension

筛选标记(Selection Marker)

Puromycin (5 μg/mL) + Hygromycin (20 μg/mL)

培养基(Complete Growth 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.

质量管理控制体系(QMS)

  1. 质量管理体系(ISO, GMP)
  2. 质量优势
  3. 质控流程
 

Receptor Assay

Fc gamma RI / CD64 FACS

Expression analysis of human CD64 on Human CD64 (Luc) Jurkat Reporter Cell by FACS.
Human CD64 (Luc) Jurkat Reporter Cell or negative control cell were stained with PE-labeled anti-human CD64 antibody.

Protocol

 

Application

Fc gamma RI / CD64 APPLICATION

ADCP response to anti-human CD20 antibody (RLU).
Anti-human CD20 antibody-induced ADCP activity was evaluated using Human CD64 (Luc) Jurkat Reporter Cell in the presence of Raji cells that express CD20 endogenously. The EC50 of anti-human CD20 antibody was approximately 0.0021 μg/mL.

Protocol

Fc gamma RI / CD64 APPLICATION

ADCP response to anti-human CD20 antibody (FOLD).
Anti-human CD20 antibody-induced ADCP activity was evaluated using Human CD64 (Luc) Jurkat Reporter Cell in the presence of Raji cells that express CD20 endogenously. The max induction fold was approximately 667.

Protocol

 

Passage Stability

Fc gamma RI / CD64 PASSAGE

Passage stability analysis by Signaling Bioassay.
The continuously growing Human CD64 (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 7-32.

Protocol

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背景(Background)

Fcγ receptors (FcγRs), the largest group of FcRs, bind IgG and comprise several subtypes. FcγRI (CD64), a high-affinity receptor that binds monomeric uncomplexed IgG molecules. FcγRI saturation by endogenous circulating IgGs in vivo may attenuate its role in mediating antibody function.

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.

  1. 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.
  2. 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.
  3. 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.
  4. 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.
  5. ACROBIOSYSTEMS ACCEPTS NO LIABILITY IN CONNECTION WITH THE HANDLING OR USE OF THE CELL LINE.
  6. 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.cn@acrobiosystems.com for further details.

 

前沿进展

Serum cytokine profiles in children with IgA vasculitis with nephritis
Jin, He, Lin et al
Biomol Biomed (2024)
Abstract: This study aimed to discover novel serum biomarkers for IgA vasculitis with nephritis (IgAVN). The serum of IgA vasculitis (IgAV) patients without nephritis and IgAVN patients not treated with glucocorticoids was analyzed for 440 proteins using a novel quantitative planar protein microarray. To verify the biomarkers, semiquantitative immunofluorescence analysis was performed on selected differential cytokines in a separate cohort of kidney tissue samples. A total of 41 proteins were differentially expressed between the IgAVN and IgAV groups out of the 440 proteins analyzed. Five differentially abundant proteins, including VEGF R3, ADAM12, TIM-3, IL-12p40, and CEACAM-5, were further validated by semiquantitative immunofluorescence analysis in kidney tissue from independent cohorts. ADAM12, TIM-3, IL-12p40, and CEACAM-5 were expressed in kidney tissue. A linear relationship was observed between the pathological grade of IgAVN and the expression levels of ADAM12 and CEACAM-5. Furthermore, while the prognosis of children with IgAVN may have a linear relationship with CEACAM-5, the results did not indicate a significant statistical difference, which may be related to the sample size. The expression of ADAM12 and CEACAM-5 was positively correlated with the pathological grade. More importantly, we found that CEACAM-5 may be related to the prognosis of IgAVN, which could serve as a significant biomarker for assessing disease severity and monitoring disease progression.
Development of a highly sensitive immunoassay based on pentameric nanobodies for carcinoembryonic antigen detection
Gu, Guo, Deng et al
Anal Chim Acta (2023) 1279, 341840
Abstract: Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM-5) is a well-characterized biomarker for the clinical diagnosis of various cancers. Nanobodies, considered the smallest antibody fragments with intact antigen-binding capacity, have gained significant attention in disease diagnosis and therapy. Due to their peculiar properties, nanobodies have become promising alternative diagnostic reagents in immunoassay. However, nanobodies-based immunoassay is still hindered by small molecular size and low antigen capture efficacy. Therefore, there is a pressing need to develop novel nanobody-based immunoassays with superior performance.A novel pentameric nanobodies-based immunoassay (PNIA) was developed with enhanced sensitivity and specificity for CEACAM-5 detection. The binding epitopes of three anti-CEACAM-5 nanobodies (Nb1, Nb2 and Nb3) were analyzed. To enhance the capture and detection efficacy of CEACAM-5 in the immunoassay, we engineered bispecific nanobodies (Nb1-Nb2-rFc) as the capture antibody, and developed the FITC-labeled pentameric nanobodies (Nb3-VT1B) as the detection antibody. The binding affinities of Nb1-Nb2-rFc (1.746 × 10-10) and Nb3-VT1B (1.279 × 10-11) were significantly higher than those of unmodified nanobodies (Nb1-rFc, 4.063 × 10-9; Nb2-rFc, 2.136 × 10-8; Nb3, 3.357 × 10-9). The PNIA showed a linear range of 0.625-160 ng mL-1 with a correlation coefficient R2 of 0.9985, and a limit of detection of 0.52 ng mL-1, which was 24-fold lower than the immunoassay using monomeric nanobody. The PNIA was validated with the spiked human serum. The average recoveries ranged from 91.8% to 102% and the coefficients of variation ranged from 0.026% to 0.082%.The advantages of nanobodies offer a promising alternative to conventional antibodies in disease diagnosis. The novel PNIA demonstrated superior sensitivity and high specificity for the detection of CEACAM-5 antigen. This bispecific or multivalent nanobody design will provide some new insights into the design of immunoassays for clinical diagnosis.Copyright © 2023 Elsevier B.V. All rights reserved.
Extracellular Vesicles Released from Cancer Cells Promote Tumorigenesis by Inducing Epithelial to Mesenchymal Transition via β-Catenin Signaling
Malyla, Paudel, Rubis et al
Int J Mol Sci (2023) 24 (4)
Abstract: Lung cancer is the leading cause of cancer-related deaths globally, in part due to a lack of early diagnostic tools and effective pharmacological interventions. Extracellular vesicles (EVs) are lipid-based membrane-bound particles released from all living cells in both physiological and pathological states. To understand the effects of lung-cancer-derived EVs on healthy cells, we isolated and characterized EVs derived from A549 lung adenocarcinoma cells and transferred them to healthy human bronchial epithelial cells (16HBe14o). We found that A549-derived EVs carry oncogenic proteins involved in the pathway of epithelial to mesenchymal transition (EMT) that are regulated by β-catenin. The exposure of 16HBe14o cells to A549-derived EVs resulted in a significant increase in cell proliferation, migration, and invasion via upregulating EMT markers such as E-Cadherin, Snail, and Vimentin and cell adhesion molecules such as CEACAM-5, ICAM-1, and VCAM-1, with concomitant downregulation of EpCAM. Our study suggests a role for cancer-cell-derived EVs to induce tumorigenesis in adjacent healthy cells by promoting EMT via β-catenin signaling.
Development and Characterization of an Anti-Cancer Monoclonal Antibody for Treatment of Human Carcinomas
Tsang, Fantini, Mavroukakis et al
Cancers (Basel) (2022) 14 (13)
Abstract: NEO-201 is an IgG1 humanized monoclonal antibody (mAb) that binds to tumor-associated variants of carcinoembryonic antigen-related cell adhesion molecule (CEACAM)-5 and CEACAM-6. NEO-201 reacts to colon, ovarian, pancreatic, non-small cell lung, head and neck, cervical, uterine and breast cancers, but is not reactive against most normal tissues. NEO-201 can kill tumor cells via antibody-dependent cell-mediated cytotoxicity (ADCC) and complement dependent cytotoxicity (CDC) to directly kill tumor cells expressing its target. We explored indirect mechanisms of its action that may enhance immune tumor killing. NEO-201 can block the interaction between CEACAM-5 expressed on tumor cells and CEACAM-1 expressed on natural killer (NK) cells to reverse CEACAM-1-dependent inhibition of NK cytotoxicity. Previous studies have demonstrated safety/tolerability in non-human primates, and in a first in human phase 1 clinical trial at the National Cancer Institute (NCI). In addition, preclinical studies have demonstrated that NEO-201 can bind to human regulatory T (Treg) cells. The specificity of NEO-201 in recognizing suppressive Treg cells provides the basis for combination cancer immunotherapy with checkpoint inhibitors targeting the PD-1/PD-L1 pathway.
Showing 1-4 of 14 papers.
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联系电话:
+86 400-682-2521(全国)
010-53681107(北京)
021-50850665(上海)
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order.cn@acrobiosystems.com
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tech.cn@acrobiosystems.com
Fc gamma RI / CD64靶点信息
英文全称:High affinity immunoglobulin gamma Fc receptor I
中文全称:高亲和力免疫球蛋白γFc受体I
种类:Homo sapiens
上市药物数量:0详情
临床药物数量:0详情
最高研发阶段:临床前
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