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Human OKT3 scFV Stable Cell Line - Raji

Human OKT3 scFV Stable Cell Line - Raji

Cat.No. :  CSC-RO0914 Host Cell:  Raji

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Cell Line Information

Cell Culture Information

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Cat. No. CSC-RO0914
Description This cell line is engineered to stably overexpress membrane OKT3 (anti-CD3 monoclonal antibody) scFV sequence.
Gene OKT3 scFV
Gene Species Homo sapiens (Human)
Host Cell Raji
Host Cell Species Homo sapiens (Human)
Stability Validated for at least 10 passages
Application

1. Studying the interactions between immune cells and cancer cells

2. Studying the mechanisms of resistance to immune checkpoint blockade

3. High-throughput screening

4. Drug target validation

Quality Control Negative for bacteria, yeast, fungi and mycoplasma.
Shipping Dry ice
Storage Liquid nitrogen
Revival Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media.
Mycoplasma Negative
Format One frozen vial containing millions of cells
Storage Liquid nitrogen
Safety Considerations

The following safety precautions should be observed.

1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.

2. No eating, drinking or smoking while handling the stable line.

3. Wash hands after handling the stable line and before leaving the lab.

4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.

5. All waste should be considered hazardous.

6. Dispose of all liquid waste after each experiment and treat with bleach.

Ship Dry ice
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Background

Case Study

Applications

Publications

Q & A

Customer Reviews

This cell line has been genetically modified to overexpress the membrane-bound single-chain variable fragment (scFV) sequence of OKT3 (anti-CD3 monoclonal antibody). Expressed on the surface of mature T lymphocytes, the CD3 molecule is a complex of peptide chains bound together by non-covalent interactions that is essential in preventing acute transplant rejection. In order to phosphorylate tyrosines (Y) in the immune receptor tyrosine-based activation motifs of CD3 molecules, the CD3 molecule binds to the T cell receptor (TCR) to form a complex receptor. Tyrosine-containing proteins downstream are further phosphorylated by pY, starting a series of kinase activation events that control target gene expression and transcription related to T cell activation and proliferation. Activation signals are transmitted and TCR structure is stabilized by the CD3 molecule. Monoclonal antibodies that specifically target CD3 can remove effector T cells, stimulate or block T cell activation signal transduction, or increase the generation of regulatory T cells. These effects open up new therapeutic options for the treatment of autoimmune disorders and organ transplant rejection.

T cells are important elements of the adaptive immune response's cellular arm. The researchers used high or low-affinity anti-CD3 single-chain variable fragments (scFv) with predetermined molecular lengths to investigate the significance of CD45 segregation from engaged T cell receptors (TCRs) in starting signaling. They found that both short and elongated high-affinity anti-CD3 scFv generated equal levels of calcium mobilization, Zap70 phosphorylation, and cytokine production in Jurkat T cells using total internal reflection microscopy on glass-supported lipid bilayers. On the other hand, extended ligands for activated TCR microclusters separated CD45 from them less than short ligands. It's interesting to note that only high-affinity elongated anti-CD3 scFv activated T cells during the early phases, despite the fact that both low and high affinity scFv caused comparable levels of CD3 co-localization with CD45.

Figure 1 caption describes the construction of lipid membrane-tethered anti-CD3 antibodies, including the retroviral cassette expressing soluble anti-CD3 scFv, schematic drawings of the ligands used (OKT3 or OKT3MA scFv elongated by an immunoglobulin-like domain or the CD43 ectodomain), and artificial antigen-presenting cells (APCs) formed from glass beads coated with a lipid bilayer decorated with anti-CD3 scFv. (doi: 10.3389/fimmu.2018.00713.)Figure 1. The scientists subcloned scFv genes onto a retro vector to create OKT3-ScFv cell lines. By inserting packaged viruses into 3T3 cells and looking for steady expression, recombinant scFv was created. Talon superflow was used to purify the soluble scFv, which was then preserved for later use. (Al-Aghbar MA, et al., 2018)

1. Gene Therapy for Transplant Rejection: Examine gene therapy approaches to avoid acute transplant rejection using the Human OKT3 scFV Stable Cell Line - Raji. It may be possible to modify T cell activation signals by studying the overexpression of OKT3's membrane-bound single-chain variable fragment (scFV) sequence, which could open up new treatment options for organ transplant recipients. 2. Autoimmune Disease Therapy: Investigate the use of monoclonal antibodies directed against CD3 in the management of autoimmune diseases. Researchers can evaluate the effectiveness of CD3-targeted monoclonal antibodies in eliminating effector T cells and controlling T cell activation signals by employing the stable cell line, which could lead to the development of new therapeutic approaches for autoimmune disorders. 3. Enhanced T Cell Activation Studies: Examine the phosphorylation of tyrosines in immune receptor tyrosine-based activation patterns of CD3 molecules to learn more about the mechanisms behind T cell activation and proliferation. Studying the downstream kinase activation processes that regulate target gene expression and transcription in relation to T cell activation and proliferation is made easier with the help of stable cell lines. 4. Regulating T Cell Generation Assess the effect of monoclonal antibodies that target CD3 on the development of regulatory T cells. Through the manipulation of T cell activation signals, scientists can investigate the possibility of stimulating regulatory T cell populations to repress immunological responses, providing new perspectives on immune modulation in diverse clinical scenarios.
Customer Q&As
How stable is this human OKT3 scFV stable cell line? Is there long-term data support?

A: Our human OKT3 scFV stable cell line has undergone long-term stability testing and maintains stable expression levels over multiple passages. We regularly validate cell lines and record cell line stability data. Customers can request this data from us to understand the stability performance of our products.

How functional is this stable cell line when studying OKT3 scFV-mediated immune responses? Is relevant functional verification data provided?

A: Our human OKT3 scFV stable cell line is fully functional and widely used in immune response research. We provide detailed functional validation data, including cell proliferation experiments, cytokine release analysis, and more. Customers can refer to these data to evaluate the performance of our products.

Is this human OKT3 scFV stable cell line suitable for in vitro cell therapy research? Is there relevant preclinical research data to support this?

A: Our human OKT3 scFV stable cell line has been validated in in vitro cell therapy studies. We provide relevant preclinical research data support to prove the application potential of our products in these fields. Customers can refer to these data to understand the performance and application scope of our products in preclinical studies.

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Customer Reviews
Grow well

This cell line exhibits highly stable gene expression during culture, particularly overexpression of the OKT3 (anti-CD3 monoclonal antibody) scFV sequence. This high-efficiency expression plays an important role in inhibiting acute transplant rejection. I am also very satisfied with the supplier's professional quality and quick response service. Both technical support and product quality have left a deep impression on me.

United States

06/24/2022

Technical Support

Recently, I started experimenting with the Human OKT3 scFV Stable Cell Line - Raji cell line and had a great experience. The cells grew well during the culture process, and their gene expression was stable and consistent, especially in the interaction between T cell receptor (TCR) and CD3 molecules. This cell line is effective in promoting T cell activation and proliferation and is suitable for a variety of immune-related research. I am also very satisfied with the platform's professional support and meticulous service. Their response speed and technical guidance are very timely and effective.

United States

03/02/2022

Excellent performance

Its efficient expression of CD3 molecules makes T cell activation and proliferation more stable, and it also performs well in downstream kinase activation events. This cell line has provided great help in my research on autoimmune diseases and organ transplant rejection. At the same time, the platform's delivery speed and after-sales service were also very good, ensuring that my experiment went smoothly.

Germany

11/23/2020

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