Recombinant anti-mouse IL-28b antibody

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 mouse IL-28B antibody

Neutralizing monoclonal antibody against murine IL-28B

InvivoGen provides a recombinant fully mouse anti-mouse IL-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 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].

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-λ 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.