Murine Anti-mIL-28b mAb

Notification: Our anti-mouse IL-28B antibody raised in mice and produced in hybridoma (mabg-mil28b/mabg-mil28b-5) has been removed from our catalog. Its coding sequence has been determined and the antibody is now produced by recombinant technology and purified from CHO cells.

Recombinant mAb against murine IL-28B

Neutralizing monoclonal antibody against murine IL-28B

InvivoGen provides a recombinant anti-mIL-28b-mIgG1 monoclonal antibody (mAb) that was previously extracted from hybridoma. It is now expressed and produced in chinese hamster ovary (CHO) cells, ensuring reliability and lot-to-lot reproducibility. Thereby, common hybridoma-related drawbacks such as the generation of non-relevant mAbs containing aberrant light chains are avoided [1].

The sequence of the anti-mIL-28b-mIgG1 is 100% murine (constant and variable regions), as the original clone (clone 3C11) was raised in mice using a proprietary method. This feature allows for reduced immunogenicity and risks of fatal hypersensitivity reactions upon repeated mAb injections into mice [2-4].

InvivoGen provides this antibody in two grades:

• In vitro use: Anti-mIL-28b-mIgG1
• In vivo use: Anti-mIL-28b-mIgG1 InvivoFit™

All InvivoFit™ products are handled in a clean room, filter-sterilized, and tested for bacterial contaminants. Additionally, this grade guarantees a low levels of endotoxins (<1 EU/ml). The buffer formulation is specifically adapted for in vivo studies.

Key features:

• Potent and specific neutralizing activity against mIL-28B
• Sequence is 100% murine
• Murine IgG1 isotype (constant region)
• Free from non-relevant mAbs found in hybridoma-based productions
• Produced in animal-free facilities and defined media
• Low aggregation < 5%
• InvivoFit™ grade is available

Anti-mIL-28b-mIgG1 is designed to efficiently neutralize the biological activity of mIL-28B. Interleukin 28B (IL-28B) is a member of the type III interferon (IFN) family,  that exhibits several common features with type I IFNs: antiviral and antitumor activity [5,6].

More details

References:

1. Bradbury, A. et al., 2018. When monoclonal antibodies are not monospecific: Hybridomas frequently express additional functional variable regions. mAbs, 10(4), 539–546.
2. Mall C. et al., 2016. Repeated PD-1/PD-L1 monoclonal antibody administration induces fatal xenogenic hypersensitivity reactions in a murine model of breast cancer. Onco Immunol. 5(2):e1075114.
3. Murphy, J.T. et al., 2014. Anaphylaxis caused by repetitive doses of a GITR agonist monoclonal antibody in mice. Blood. 123(14):2172-2180.
4. Belmar N.A. et al., 2017. Murinization and H chain isotype matching of Anti-GITR antibody DTA-1 reduces immunogenicity-mediated anaphylaxis in C57BL/6 mice. J Immunol. 198:4502-4512.
5. Donnelly RP. et al., 2010. Interferon-lambda: a new addition to an old family. J Interferon Cytokine Res. 30(8):555-64.
6. Li M. et al., 2009. Interferon-ls: the modulators of antivirus, antitumor, and immune responses. J. Leukoc. Biol., 86:23-32.

Figures

Anti-mIL-28b-mIgG1 InvivoFit™ is a potent neutralization antibody. Increasing concentrations of Anti-mIL-28b-mIgG1 InvivoFit™ were incubated for 30 minutes with the recombinant cytokine mIL28B (30 ng/ml) prior to the addition of HEK-Blue™ IFN-α/β cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown in percentage (%) of neutralization.

Specifications

Target: Murine  IL-28B (mIL-28B)

Specificity: Reacts with murine IL-28A and murine IL-28B
No cross-reactivity with human IL-28A or human IL-28B

Clone: 3C11

Source: CHO cells

Isotype: Murine IgG1, kappa 

Control: Murine IgG1 Isotype Control

Formulation of Anti-mIL-28b-mIgG1: Lyophilized from 0.2 µm filtered solution in a sodium phosphate buffer with glycine, saccharose, and stabilizing agents

Formulation of Anti-mIL-28b-mIgG1 InvivoFit™: Lyophilized from 0.2 µm filtered solution in a sodium phosphate buffer with 5% saccharose

Tested applications: Blocking & Neutralization

Quality control:

• These products have been validated using neutralization cellular assays.
• The complete sequence of these antibodies has been verified.
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK‑Blue™ TLR4 cells.
• The endotoxin level in Anti-mIL-28b-mIgG1 InvivoFit™ is <1 EU/mg (determined by a LAL assay).

Contents

Please note: each mAb is sold separately.

Anti-mIL-28b-mIgG1

• mil28b-mab9-02​: 200 µg, lyophilized

Anti-mIL-28b-mIgG1 InvivoFit™

• mil28b-mab9-1​: 1 mg, lyophilized

 The product is shipped at room temperature.

 Store lyophilized antibody at -20 °C.

 Lyophilized product is stable for at least 1 year.

 Avoid repeated freeze-thaw cycles.

InvivoFit™

InvivoFit™ is a high-quality standard specifically adapted to in vivo studies. InvivoFit™ products are filter-sterilized (0.2 µm) and filled under strict aseptic conditions in a clean room. The level of bacterial contaminants (endotoxins and lipoproteins) in each lot is verified using a LAL assay and a TLR2 and TLR4 reporter assay.

Details

Type III interferons (IFNs) - also called IFN-λ cytokines - play an essential role within the antiviral response in epithelial barrier tissues. In humans, this family comprises four distinct proteins called IFN-λ1 (IL-29), IFN-λ2 (IL-28A), IFN-λ3 (IL-28B), and the poorly secreted IFN-λ4. In mice, two functional orthologs (IL-28A and IL-28B) have been described [1,2]. IFN-λs are produced by virtually any nucleated cell type. However, the target cell populations of IL-28/IL-29 are restricted and mainly include epithelial cells and hepatocytes [3].

Upon infection, viral RNA is sensed by pattern-recognition receptors (PRRs) including RIG-I and MDA5 [3]. This stimulates the transcription of IFN-λs by IRF1 (IFN regulatory factor 1). Subsequently, IFN-λs bind to a heterodimeric receptor formed by the assembly of IFNLR1 (IFN-lambda receptor 1) and IL10Rβ (IL-10 receptor β). This leads to the recruitment of the Janus kinases, JAK1 and TyK2, and phosphorylation of STAT1 and STAT2. They dimerize and interact with IRF9, forming the so-called ISGF3 complex. ISGF3 translocates into the nucleus and binds to IFN-stimulated response elements (ISRE) within the promoter region of antiviral IFN-stimulated genes (ISG) to regulate their expression [2-3]. Besides its antiviral function, IFN-λ signaling plays also an important role in non-infectious diseases like inflammatory bowel disease and cancer [2].

1. Lazear HM. et al., 2015. Interferon-λ: Immune Functions at Barrier Surfaces and Beyond. Immunity. 43(1):15-28.
2. Witte K.et al., (2010). IL-28A, IL-28B, and IL-29: Promising cytokines with type I interferon-like properties. 21(4), 0–251.
3. Lee S. & Baldridge MT., 2017. Interferon- Lambda: A Potent Regulator of Intestinal Viral Infections. Front Immunol. 8:749.