Anti-CD20-Anti-CD3 IgM format Bispecific Antibody

Only %1 left

Catalog #

100860

As low as $390 *

Cat # Size Price*

Qty

Bulk/Custom

Add to Favorites

*US Pricing only. For international pricing, please contact your local distributor.

●

Purchase

●

Description

His-tagged Anti-CD20-Anti-CD3 IgM format bispecific antibody is a purified recombinant human bispecific antibody with T cell Engager. This bispecific antibody has been tested for specific activity in the functional reporter assay using NFAT-luc reporter Jurkat cell line (BPS Bioscience #60621) in the presence of CD20-CHO cells (BPS Bioscience #79624-H). The IgM Fc region is pentameric, so there are 5 copies of the anti-CD20 antibodies and J chain has anti-CD3 antibody.

●

Synonyms

Anti-CD20 BiTE®, Anti-CD3 BiTE®, IgM bispecific antibody

●

Product Data Gallery

4-20% SDS-Page Coomassie Staining

Activation of NFAT-Jurkat Reporter by AntiCD20-Anti-CD3 IgM

Product Info

Storage and Usage

Citations

Species

Human

Construct

Anti-CD20-IgM(1-594(end)-His, 1-241(end)) / Anti-CD3-Jchain(1-415(end))

Isotype

IgM

Host Species/Expression System

HEK293

Purity

≥90%

Supplied As

Aqueous buffer solution.

Purification

Ni-NTA affinity purification of the His-tag protein from HEK293 cells.

Formulation

8 mM phosphate, 110 mM NaCl, 2.2 mM KCl, pH 7.4, and 20% glycerol

Anti-CD20 HC: 66 kDa; Anti-CD20 LC: 26 kDa; Anti-CD3 VC: 45 kDa

Amino Acids

Anti-CD20 HC: 1-594(end); Anti-CD20 LC: 1-241(end); Anti-CD3 VC: 1-415(end)

UniProt #

P11836

Tag(s)

Anti-CD20: C-terminal His-Tag

Background

CD20 (MS4A1) is a glycosylated phosphoprotein expressed on the cell surface of B cells. Although the functional significance of CD20 is not clear, and CD20 has no known ligands, CD20 has been shown to regulate intracellular calcium levels. CD20 is a highly attractive target antigen for immunotherapy because it is expressed in more than 90% of patients with B-cell lymphoma. First approved in 1997, Rituximab (Rituxan) is a chimeric monoclonal antibody targeting CD20 and has been classified by the World Health Organization as an “Essential Medicine”. Since then, additional monoclonal antibodies against CD20 have been approved or are being tested in clinical trials for the treatment of multiple sclerosis (MS), chronic lymphocytic leukemia (CLL), follicular lymphoma, diffuse large B cell lymphoma (DLBCL), rheumatoid arthritis, non-Hodgkin’s lymphoma, systemic lupus erythematosus, and myalgic encephalomyelitis (chronic fatigue syndrome).

The IgM pentameric structure is supposed to produce a stronger binding to cells with low expression of CD20, as well as greater ability to cross-link cell surface receptors for increased cellular signaling. In the meantime, because of the ratio of anti CD20- and anti CD3-antibodies, the cytokine release is supposed to be reduced.

Storage/Stability

At least 12 months at -80°C. Avoid freeze/thaw cycles.

Assay Conditions

Experimental design and assay protocol for measuring anti CD20-anti CD3 functional activity using NFAT-luc reporter Jurkat cell line:
Jurkat effector cells with endogenous TCR/CD3 and transfected reporter NFAT-luc (BPS Bioscience #60621) are incubated with increasing concentrations of anti CD20 x anti CD3 bispecific antibody in the presence of CD20-CHO cells (BPS Bioscience #79624-H) or CHO cells (ATCC #CCL-61™).
1. Seed CHO and CD20-CHO cells at 30,000 cells/well and allow a few hours for the cells to attach in a 96-well clear bottom white plate.
2. Seed Jurkat cells at 30,000 cells/well.
3. Add the bispecific antibody at a recommended dilution range of 100 fM-100 nM. The bispecific antibody simultaneously binds to TCR/CD3 on the NFAT-luc Jurkat reporter cells and tumor antigen CD20 on CD20-CHO cells.
4. After 16 hours the luciferase activity is measured using ONE-Step™ luciferase assay (BPS Bioscience #60690) per recommended protocol. The bispecific antibody interaction stimulates NFAT-luciferase activity.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles.

Scientific Category

Immunotherapy, BiTE®

Regulatory Status

Research Use Only