LAG-3 Antibody - IgG4SP, (Research Grade relatlimab Biosimilar) - Low Endotoxin, Azide and BSA Free

Novus Biologicals | Catalog # NBP3-28028

Recombinant Monoclonal Antibody
Novus Biologicals

Key Product Details

Species Reactivity

Human

Applications

ELISA, Flow Cytometry, Functional

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Human IgG4 expressed in CHO

Format

Low Endotoxin, Azide and BSA Free
View all LAG-3 Primary Antibodies »

Product Specifications

Immunogen

LAG3 / CD223

Reactivity Notes

Predicted species reactivity: Cynomolgus

Clonality

Monoclonal

Host

Human

Isotype

IgG4

Description

The heavy chain type is huIgG4SP, and the light chain type is hukappa. It has a predicted MW of 145.5 kDa.

Also known as 'relatlimab'.

IgG4SP is human IgG4 with S228P mutation.

This product is shipped at ambient temperature. Upon receipt, store immediately at -20C or lower for 12 months in a lyophilized state. - 80C for 3 months after reconstitution. Avoid repeated freeze-thaw cycles.

Endotoxin Level

< 0.001EU/ug,determined by LAL method.

Scientific Data Images

LAG-3 Antibody (relatlimab) - IgG4SP

In vivo assay: LAG-3 Antibody (relatlimab) - IgG4SP[NBP3-28028] -

Relatlimab inhibited the tumor growth of MC38 on Human LAG3 mice. The result showed significant anti-tumor effects, with an tumor inhibition rate (TGI) of 53.8% at 10 mpk at D28.
LAG-3 Antibody (relatlimab) - IgG4SP

ELISA: LAG-3 Antibody (relatlimab) - IgG4SP[NBP3-28028] -

Immobilized human LAG3 His at 2 ug/mL can bind LAG-3 Antibody (relatlimab) - IgG4SP, EC50=0.02812 ug/mL.
LAG-3 Antibody (relatlimab) - IgG4SP

Flow Cytometry: LAG-3 Antibody (relatlimab) - IgG4SP[NBP3-28028] -

Human LAG3 CHO cells were stained with LAG-3 Antibody (relatlimab) - IgG4SP and negative control protein respectively, washed and then followed by PE and analyzed with FACS, EC50=0.5464 ug/mL.

Applications

Application
Recommended Usage

ELISA

Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Functional

Optimal dilutions of this antibody should be experimentally determined.

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A purified

Reconstitution

Reconstitute with sterile, distilled water to a final concentration of 1 mg/ml. Gently shake to solubilize completely. Do not vortex.

Formulation

Lyophilized from 25mM histidine, 8% sucrose, 0.01% Tween80 (pH6.2)

Format

Low Endotoxin, Azide and BSA Free

Preservative

No Preservative

Concentration

LYOPH mg/ml

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C in powder form. Store at -80C once reconstituted.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: LAG-3

Lymphocyte activation gene-3 (LAG-3), also referred to as CD233, is a type I transmembrane protein with a theoretical molecular weight of 70 kDa that is a member of the immunoglobulin superfamily (IgSF) (1, 2). Human LAG-3 cDNA encodes 525 amino acids (aa) that includes a 28 aa signal sequence, a 422 aa extracellular domain (ECD) with four Ig-like domains (D1-D4), a transmembrane region and a highly charged cytoplasmic region. Within the ECD, human LAG-3 shares 70%, 67%, 76%, and 73% aa sequence identity with mouse, rat, porcine, and bovine LAG-3, respectively. The extracellular region of LAG-3 and the CD4 co-receptor share ~20% aa sequence homology but are structurally similar and both bind to major histocompatibility complex class II (MHCII) on antigen-presenting cells (APCs), although LAG-3 has much higher affinity (1, 3). LAG-3 is highly expressed in the lymph node, spleen, ovary, and appendix while expressed at a lower level in a variety of other tissues. More specifically, LAG-3 is expressed on activated CD4+ and CD8+ T cells, natural killer T (NKT) cells, natural killer (NK) cells, plasmacytoid dendritic cells (pDCs), and regulatory T cells (Tregs), but not on naive, or resting, T cells (1, 3).

As mentioned above, LAG-3 binds to MHCII and this occurs via a proline-rich amino acid loop in D1 (1, 3). Another unique feature of LAG-3 is the longer connecting peptide region between the D4 and the transmembrane, which is acted upon and cleaved by metalloproteinases a disintegrin and metallopeptidase domain (ADAM) 10 and ADAM17 to generate a soluble 54 kDa form of LAG-3 (sLAG-3) (1, 3). The interaction of LAG-3 with MHCII prevents the MHC molecule from binding to a T-cell receptor (TCR) or CD4, thereby functioning in an inhibitory role and suppressing the TCR signal (4). When LAG-3 crosslinks with the TCR/CD3 complex, it causes reduced T-cell proliferation and cytokine secretion (4). This negative regulation is important in controlling autoimmunity as one study found Lag3-/- NOD (non-obese diabetic) mice had accelerated diabetes onset and increased T-cell infiltration into islet cells (5). On the other hand, besides being a negative regulator of T-cells, LAG-3 binding to MHCII molecules on APCs induces dendritic cell maturation and cytokine secretion by monocytes (5, 6). In addition to MHCII, other reported ligands for LAG-3 includes fibrinogen-like protein 1 (FGL1), liver endothelial cell lectin (lSECtin), galectin-3 (Gal-3), and alpha-synuclein fibrils (1). Gal-3, for instance, is expressed on stromal cells and CD8+ T-cells in the tumor microenvironment and the interaction with LAG-3 was shown to be crucial for the suppression of secreted cytokine IFN-gamma and may control anti-tumor immune responses (1, 5). Interestingly, a mouse model of Parkinson's disease revealed LAG-3 binding to alpha-synuclein fibrils in the central nervous system, contributing to its pathogenesis (1, 5).

Recent cancer immunotherapeutic approaches have focused on inhibitory receptors such as LAG-3 to revive expression of cytotoxic T-cells to attack tumors (6). LAG-3 has been shown to be co-expressed and have synergy with another immune-checkpoint molecule called programmed-death 1 (PD-1) (1, 4, 5, 6). In a mouse model of colon adenocarcinoma LAG3 blockade alone was largely ineffective, however co-blockade of LAG-3 and PD-1 limited tumor growth and resulted in tumor clearance in 80% of mice, compared to 40% with PD-1 blockade alone (5). Additionally, in a model of fibrosarcoma the LAG-3/PD-1 duel blockade increased survival and the percentage of tumor-free mice (5). Analysis of a variety of human tumor samples (e.g. melanoma, colon cancer, head and neck squamous cell carcinoma) also suggest that LAG3 alone and combinatorial treatment with PD-1 may be a good target for treatment (1, 3-6). To date there are over 10 different agents targeting LAG-3 in clinical trials for cancer either as an anti-LAG-3 blocking antibody monotherapy or as a combination antagonist bispecific antibody, primarily with PD-1 (1, 3-6).

Alternative names for LAG-3 includes 17b4 lag3, 17b4 neutralizing, 17b4, CD223, FDC, LAG-3 17b4, LAG-3 blocking, and LAG3.

References

1. Maruhashi, T., Sugiura, D., Okazaki, I. M., & Okazaki, T. (2020). LAG-3: from molecular functions to clinical applications. Journal for Immunotherapy of Cancer, 8(2), e001014. https://doi.org/10.1136/jitc-2020-001014

2. Triebel, F., Jitsukawa, S., Baixeras, E., Roman-Roman, S., Genevee, C., Viegas-Pequignot, E., & Hercend, T. (1990). LAG-3, a novel lymphocyte activation gene closely related to CD4. The Journal of experimental medicine, 171(5), 1393-1405. https://doi.org/10.1084/jem.171.5.1393

3. Ruffo, E., Wu, R. C., Bruno, T. C., Workman, C. J., & Vignali, D. (2019). Lymphocyte-activation gene 3 (LAG3): The next immune checkpoint receptor. Seminars in immunology, 42, 101305. https://doi.org/10.1016/j.smim.2019.101305

4. Long, L., Zhang, X., Chen, F., Pan, Q., Phiphatwatchara, P., Zeng, Y., & Chen, H. (2018). The promising immune checkpoint LAG-3: from tumor microenvironment to cancer immunotherapy. Genes & cancer, 9(5-6), 176-189.

5. Andrews, L. P., Marciscano, A. E., Drake, C. G., & Vignali, D. A. (2017). LAG3 (CD223) as a cancer immunotherapy target. Immunological reviews, 276(1), 80-96. https://doi.org/10.1111/imr.12519

6. Goldberg, M. V., & Drake, C. G. (2011). LAG-3 in Cancer Immunotherapy. Current topics in microbiology and immunology, 344, 269-278. https://doi.org/10.1007/82_2010_114

Long Name

Lymphocyte-activation Gene 3

Alternate Names

CD223, LAG3

Gene Symbol

LAG3

UniProt

P18627

Additional LAG-3 Products

Product Documents

Certificate of Analysis

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Product Specific Notices

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

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