Anti-Mouse 4-1BB Monoclonal Antibodies

PA007275.r1: In vivo Grade Recombinant Anti-mouse CD137 (TNFRSF9 or 4-1BB) Monoclonal Antibody, Rat IgG1 Kappa (Clone: LOB12.3)

Recombinant rat IgG1 Monoclonal Antibody. 
Clone: LOB12.3.
Isotype: Rat IgG1 kappa.
Source: The anti-mouse CD137 (TNFRSF9 or 4-1BB) monoclonal antibody (clone: LOB12.3) was produced in mammalian cells.
Specificity/Sensitivity: The in vivo grade recombinant rat monoclonal antibody (clone: LOB12.3) specifically binds to the mouse 4-1BB protein.
Applications: ELISA, flow cytometry, neutralization, functional assays such as bioanalytical PK and ADA assays, and those assays for studying biological pathways affected by the mouse 4-1BB protein. 
Form of Antibody: 0.2 uM filtered solution, pH 7.4, no stabilizers or preservatives.
Endotoxin: < 1 EU per 1 mg of the protein by the LAL method.
Purity: >95% by SDS-PAGE under reducing conditions and HPLC.

Shipping: The antibody is shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70°C as supplied.
1 month from date of receipt, 2 to 8°C as supplied.

Background

The LOB12.3 antibody binds to 4-1BB (TNFRSF9 or CD137), one representative TNF receptor family co-stimulatory receptor. The 4-1BB protein is expressed on a wide variety of cell types, including activated T cells, NK cells, DCs, B cells, monocytes, and neutrophils. Anti-4-1BB-induced CD8+ T responses play a dominant role in anti-tumor immunity, such as induction of more effector molecules released from CD8+ T cells, increased proliferation and decreased apoptosis of CD8+ T cells.

References of Anti-mouse CD137 (TNFRSF9 or 4-1BB) Monoclonal Antibody:

Optimization of 4-1BB antibody for cancer immunotherapy by balancing agonistic strength with FcγR affinity.
Qi, X., et al. Nat Commun. 2019 May 20;10(1):2141. doi: 10.1038/s41467-019-10088-1. PMID: 31105267
To generate anti-human 4-1BB antibody LYGN6051, Balb/C and SJL mice were immunized with human 4-1BB protein and pcDNA3.1-human CD137 plasmid. ...Both LOB12.3 and 3H3 Abs showed anti-tumor efficacy (Fig. 1a, b) but they exhibited distinct liver toxicity profiles; 3H3 significantly increased alanine transaminase (ALT) levels, whereas LOB12.3 had minimal impact on ALT levels. ...Surprisingly, only 3H3 showed a co-stimulatory effect, suggesting the co-stimulatory ability of 3H3 does not depend on non-T cells while that of LOB12.3 requires help from other cells. ...In co-cultures of FcγRIIB-expressing or FcγRIII-expressing cells and purified CD8+ T cells, LOB12.3’s co-stimulatory activity was restored. ...Both 3H3-rIgG2a and LOB12.3-rIgG1 showed a protective effect, suggesting an existing tumor-specific memory response (Supplementary Fig. 4F).
Tags: anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3; anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 antibody

Curing mice with large tumors by locally delivering combinations of immunomodulatory antibodies.
Dai, M., et al. Clin Cancer Res. 2015 Mar 1;21(5):1127-38. doi: 10.1158/1078-0432.CCR-14-1339. PMID: 25142145
More than 50% of tumor-bearing mice had complete regression and long-term survival after tumor injection with mAbs recognizing CD137/PD-1/CTLA-4/CD19 with similar responses in three models. ...Intratumoral injection of mAbs recognizing CD137/PD-1/CTLA-4/CD19 can eradicate established tumors and reverse a Th2 response with tumor-associated CD19 cells to Th1 immunity, whereas a combination lacking anti-CD19 is less effective. ...On the basis of these findings and our previous demonstration that a combination of mAbs to CD137/PD-1/CTLA-4 prolongs survival of mice with the ID8 ovarian carcinoma (35, 36), as well as data from others (as cited above), we conclude that many cancers can be cured by engaging immunologic mechanisms of the Th1 type and we expect that increased knowledge about the underlying mechanisms will revolutionize cancer therapy. ...The following mAbs were purchased from BioXcell: anti-CD137 (LOB12.3; Cat. #BE0169), anti–PD-1 (RMP1-14; Cat. #BE0146), anti–CTLA-4 (9D9; Cat. #BE0164), anti-CD19 (1D3; Cat. #BE0150), and control (2A3; Cat. #BE0089), and administered as indicated. ...It also increased the number of cells expressing CD86 or CD137, while expression of PD-1 and CTLA-4 was unaffected in both TLN and spleen (Supplementary Fig. S2).
Tags: anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 in vivo; anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 mAb in vivo

Anaphylaxis caused by repetitive doses of a GITR agonist monoclonal antibody in mice.
Murphy, J. T., et al. Blood. 2014 Apr 3;123(14):2172-80. doi: 10.1182/blood-2013-12-544742. PMID: 24558202
DTA-1 (anti-GITR), OX86 (anti-OX40), FGK45.5 (anti-CD40), GK1.5 (anti-CD4), 11B11 (anti-IL-4), and TA99 (anti-Tyrp-1) were produced and purified by the Monoclonal Antibody Core Facility at MSKCC. LOB12.3 (anti-4-1BB), 1A8 (anti-Ly6G), LTF-2 (rat immunoglobulin G2b [IgG2b] isotype control), and HRPN (rat IgG1 isotype control) were purchased from Bio X Cell. ...For experiments investigating anaphylaxis, 1 mg of DTA-1, OX86, FGK45.5, LOB12.3, LTF-2, or OVA was injected on days 0, 4, and 11. For IL-4 neutralization experiments, 500 µg 11B11 was injected on day 0 of DTA-1 treatment. ...All reagents were purchased from Thermo Scientific unless otherwise stated. ...A disposable 5-mL polypropylene column was packed with 1 mL of Protein A/G agarose. ...Mice were treated with DTA-1 or LTF-2 on days −11 and −7 relative to sera collection. Sera were analyzed by enzyme-linked immunosorbent assay by using plates coated with either DTA-1 or LTF-2 in PBS and biotinylated rat anti-mouse IgG1 (BD Biosciences) as a secondary antibody.
Tags: anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 antibody in animal model; anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 in cancer research

Localized immunotherapy via liposome-anchored Anti-CD137 + IL-2 prevents lethal toxicity and elicits local and systemic antitumor immunity.
Kwong, B., et al. Cancer Res. 2013 Mar 1;73(5):1547-58. doi: 10.1158/0008-5472.CAN-12-3343. PMID: 23436794
Similarly, agonistic antibodies against the costimulatory receptor CD137 (4-1BB) expressed on the surface of activated T cells, NK cells, and dendritic cells potently inhibit tumor progression in preclinical mouse models and have begun testing in clinical trials. ...Intravenous anti-CD137 elicits liver inflammatory damage, systemic inflammatory cytokine release, hematologic abnormalities, and disruptions in lymphocyte trafficking (4, 17, 18), whereas IL-2 therapy is accompanied by potentially lethal side effects such as capillary leak syndrome. ...We hypothesized that anti-CD137 and IL-2 co-delivered via this liposome-anchored approach would synergistically activate tumor-specific T lymphocytes in the tumor and tumor-draining lymph nodes (TDLN), thereby priming effective local immune responses and the systemic dissemination of CTLs capable of targeting distal untreated lesions. ...Using the murine B16F10 model, we show here that local therapy with anti-CD137 liposomes and IL-2 liposomes leads to potent antitumor activity with no evidence for systemic toxicity, unlike i.t. soluble anti-CD137 + IL-2 treatment. ...Anti-CD137 and IL-2Fc were treated with 1.8 mmol/L dithiothreitol to expose hinge region thiols and then mixed with liposomes for covalent maleimide–thiol conjugation.
Tags: anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 mAb in cancer research; anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 antibody in mouse tumor model

Combinatorial PD-1 blockade and CD137 activation has therapeutic efficacy in murine cancer models and synergizes with cisplatin.
Wei, H., et al. PLoS One. 2013 Dec 19;8(12):e84927. doi: 10.1371/journal.pone.0084927. PMID: 24367702
In spite of the promising antitumor efficacy of several mAbs, many tumors are refractory to treatment with single anti-CD137, anti-PD-1 or anti-CTLA4 mAbs [25,26] and combinations of two or more mAbs may be needed. ...Based on these findings we have now compared the in vivo therapeutic efficacy, measured as prolonged overall survival, of anti-CD137/PD-1 combination with that of the mAbs given as single agents as well as with other mAbs and mAb combinations. ...We further investigated the systemic and local immunological mechanisms engaged by single or combined anti-CD137/PD-1 mAbs. Importantly, we subsequently showed that a combination of anti-CD137/PD-1 mAbs with the anti-cancer drug cisplatin significantly prolongs life and is probably curative to 80% of mice with established ID8 carcinoma by a mechanism that involves functional CD8+ T cells and has tumor antigen specificity as well as immunological memory. ...Mice which had been transplanted i.p. with ID8 cells were euthanized 7 and 14 days after they had been injected with the anti-PD1, anti-CD137, anti-PD1/CD137 or control as in the therapy experiments. ...Consistent with our recently published results [27], combined treatment of anti-CTLA4, PD-1 and CD137 mAbs significantly prolonged survival of mice with the mean survival time increasing to 80 days (Figure 1A,B; p<0.05 compared to controls).
Tags: function of anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3; function of anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 antibody

Combined TIM-3 blockade and CD137 activation affords the long-term protection in a murine model of ovarian cancer.
Guo, Z., et al. J Transl Med. 2013 Sep 17:11:215. doi: 10.1186/1479-5876-11-215. PMID: 24044888
Mice with established ID8 tumor were intraperitoneally injected with single or combined anti-TIM-3/CD137 monoclonal antibody (mAb); mice survival was recorded, the composition and gene expression of tumor-infiltrating immune cells in these mice was analyzed by flow cytometry and quantitative RT-PCR respectively, and the function of CD8+ cells was evaluated by ELISA and cytotoxicity assay. ...T-cell immunoglobulin and mucin domain 3 (TIM-3) is known as a negative immune regulator and emerging data have implicated TIM-3 a pivotal role in suppressing antitumor immunity. The co-stimulatory receptor CD137 is transiently upregulated on T-cells following activation and increases their proliferation and survival when engaged. ...Although antagonistic anti-TIM-3 or agonistic anti-CD137 antibodies can promote the rejection of several murine tumors, some poorly immunogenic tumors were refractory to this treatment. ...In this study, we sought to evaluate whether combined TIM-3 blockade and CD137 activation would significantly improve the immunotherapy in the murine ID8 ovarian cancer model. ...Either anti-TIM-3 or CD137 mAb alone, although effective in 3 days established tumor, was unable to prevent tumor progression in mice bearing 10 days established tumor, however, combined anti-TIM-3/CD137 mAb significantly inhibited the growth of these tumors with 60% of mice tumor free 90 days after tumor inoculation.
Tags: bioactivity of anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3; anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 antibody of low endotoxin

CD4+ T cells develop antiretroviral cytotoxic activity in the absence of regulatory T cells and CD8+ T cells.
Manzke, N., et al. J Virol. 2013 Jun;87(11):6306-13. doi: 10.1128/JVI.00432-13. PMID: 23536666
Costimulatory receptors such as CD134 and CD137 (54) induce this transcription factor after binding of their ligands. ...To provide some evidence for this hypothesis, we performed preliminary in vivo costimulation experiments with a signaling antibody against CD137. ...To characterize the role of the costimulatory receptor CD137, infected mice were treated with 100 μg of anti-CD137 (clone LOB12.3; Bioxcell) administered every other day from day four after FV infection by i.p. injection. ...The first aim of the current study was to determine the kinetics of the total CD4+ T cell response after FV infection. ...No significant increase was observed for the Th2-like cytokine IL-4. Activated CD4+ T cells also produced significantly more granzyme B (GzmB) (Fig. 2B) and showed elevated levels of perforin mRNA (data not shown), molecules that can induce apoptosis in infected target cells.
Tags: anti-mouse CD137 (TNFRSF9 or 4-1BB) LOB12.3 mAb of low endotoxin; anti-mouse CD137 (TNFRSF9 or 4-1BB) (LOB12.3) in vivo antibody

Combination CTLA-4 blockade and 4-1BB activation enhances tumor rejection by increasing T-cell infiltration, proliferation, and cytokine production.
Curran, M. A., et al. PLoS One. 2011 Apr 29;6(4):e19499. doi: 10.1371/journal.pone.0019499. PMID: 21559358
4-1BB (CD137) belongs to the Tumor Necrosis Factor Receptor (TNFR) superfamily and is transiently upregulated on both CD4+ and CD8+ T-cells following activation. ...The co-inhibitory receptor Cytotoxic T-Lymphocyte Antigen 4 (CTLA-4) is induced on T-cells shortly after activation and functions to attenuate their proliferation, IL-2 production, and contact time with antigen presenting cells (APC). ...We hypothesized that increasing the tumor-specific T-cell frequency through vaccination would allow us to better observe the interaction between 4-1BB activation and CTLA-4 blockade in the B16 melanoma system. ...To understand the apparent synergy between CTLA-4 blockade and 4-1BB activation in the context of our Flt3-ligand base vaccine, we sought to dissect the effects of each therapy on T-cell infiltration of tumor in this background. ...The failure of GVAX to support cooperativity between CTLA-4 blockade and 4-1BB activation does not appear to result from an inability to generate advantageous effector to Treg ratios.
Tags: clone LOB12.3 of anti-mouse CD137 (TNFRSF9 or 4-1BB) ; clone LOB12.3 of mouse CD137 (TNFRSF9 or 4-1BB) antibody

For more references about Anti-mouse CD137 (TNFRSF9 or 4-1BB) Monoclonal Antibody please contact our scientific support team with message@sydlabs.com.

Related Recombinant IgG Reference Antibodies:
Recombinant rat lgG1 isotype control antibody
Recombinant human IgG1 isotype control antibodies

Syd Labs provides the following anti-mouse 4-1BB antibodies:
Anti-mouse 4-1BB monoclonal antibody (Clone: 3H3)