Since the early 1960s, immunologists like Adler and colleagues have investigated the biphasic immune response—initially marked by an early IgM release, followed by a delayed but specific IgG response in immunized animals. One significant discovery emerged from studies involving T2 bacteriophage-infected peritoneal exudate cells, where immune responses in lymph nodes of naïve animals were triggered upon co-culture with these cells. Interestingly, the IgG produced reflected the donor lymph node origin, while IgM was traced back to donor exudate cells.

This led to a pivotal question in immunology: could a messenger molecule exist that facilitates communication between distinct immune cell subsets?

Macrophages within the exudate cells—responsible for phagocytosing the T2 bacteriophage—did not produce immunoglobulins. However, they appeared to influence the antibody production of other immune cells. By the late 1970s, multiple research groups hypothesized that macrophages contributed to immune regulation by processing antigens, presenting them to lymphocytes, or even transferring genetic instructions for antibody production.

In 1972, Gery and colleagues introduced the term lymphocyte-activating factor (LAF) after discovering that macrophages released substances that enhanced T-cell responses. Meanwhile, Elisha Atkins studied the fever-inducing “endogenous pyrogen” produced by leukocytes during infections. Charles Dinarello later demonstrated that LAF and endogenous pyrogen were in fact the same molecule—now known as interleukin-1 (IL-1).

In 1985, researchers including March et al. identified two distinct forms of IL-1: IL-1α and IL-1β. IL-1β, a 269-amino acid cytokine with a molecular weight of 30.7 kDa, plays a critical role in inflammation, cell proliferation, differentiation, and apoptosis. Synthesized as pro-IL-1β, it becomes biologically active (17.4 kDa) after cleavage by caspase-1, a protease within the NLRP3 inflammasome.

Mutations in the NLRP3 gene can lead to spontaneous activation of this inflammasome, causing excessive IL-1β secretion. This mechanism is central to a group of autoinflammatory diseases like Cryopyrin-Associated Periodic Syndromes (CAPS), gout, and DIRA syndrome.

Beyond these conditions, IL-1β has been implicated in a range of chronic diseases, including:

• Type 2 diabetes

• Rheumatoid arthritis

• Liver fibrosis

• Various cancers

• COVID-19

This wide-ranging impact has made IL-1β a prime target for monoclonal antibody therapy.

Interleukin-1 beta (IL-1β) is a central mediator of inflammation and plays a crucial role in both acute and chronic immune responses. Its involvement in diseases ranging from auto-inflammatory syndromes to metabolic disorders, cancer, and infectious diseases such as COVID-19, has made it a key biomarker of inflammatory activity.

Monitoring IL-1β levels is therefore critical in:

• Preclinical research studying inflammation pathways

• Drug development targeting cytokine signaling

• Immuno-oncology studies evaluating the tumor microenvironment

• Biomarker discovery in translational and systems immunology

To support these efforts, research-use-only (RUO) monoclonal antibodies against IL-1β are widely used in applications such as ELISA, flow cytometry, Western blot, and immunohistochemistry (IHC). However, producing robust and specific RUO antibodies against IL-1β remains a significant scientific challenge.

IL-1β is typically expressed at low levels under homeostatic conditions and only produced in response to specific inflammatory triggers. Even then, it is often retained in the cell in its inactive precursor form (pro-IL-1β) and only cleaved into its mature, active form by caspase-1 within the NLRP3 inflammasome.

This makes it difficult to obtain:

• Sufficient quantities of native IL-1β for immunization

• A consistent source of the active form to generate specific antibodies

IL-1β shares significant structural homology with other cytokines in the IL-1 family, including IL-1α, IL-33, and IL-18. Antibodies must be highly specific to IL-1β to avoid cross-reactivity, particularly in multiplex assays or in complex biological samples.

Generating antibodies with high specificity and affinity against IL-1β, while avoiding off-target binding, requires sophisticated screening and validation techniques, mastered by SYnAbs for many years.

IL-1β undergoes a major conformational change between its pro-form and mature form. The mature, bioactive IL-1β is the clinically relevant target, yet most epitopes are conformational and not linear, making it difficult to raise monoclonal antibodies using short peptide antigens.

SYnAbs has so been compelled to:

• Design full-length or properly folded recombinant proteins,

• Develop unique technologies with conformational peptides

• Use hybridoma multiple screening against close analogs,

• Validate antibodies for both form-specificity and species cross-reactivity

Mouse, rat-LOU, and human IL-1β differ in amino acid sequence, and so monoclonal antibodies often lack cross-species reactivity. This is problematic in translational studies where IL-1β must be monitored in animal models and human samples simultaneously.

SYnAbs therefore had to carefully design and test its monoclonal antibodies to :

• Species selectivity

• Epitope conservation

• Cross-reactivity panels

Despite these challenges, RUO monoclonal antibodies targeting IL-1β remain indispensable tools in biomedical research and reliable references on the market are rare. SYnAbs antibodies enable:

• Precise quantification of IL-1β levels in cell supernatants and serum

• Visualization of IL-1β expression in tissues under inflammatory conditions

• Mechanistic studies on inflammasome activation

• Assessment of pharmacodynamics in preclinical drug testing

Validated RUO antibodies also serve as the foundation for eventual clinical assay development and even therapeutic antibody design.

Canakinumab, a fully human monoclonal antibody, specifically targets IL-1β and inhibits its binding to the IL-1 receptor. This prevents downstream inflammatory signaling, lowering levels of IL-6, C-reactive protein (CRP), and fibrinogen. Developed by Novartis, Canakinumab received FDA approval for Cryopyrin-Associated Periodic Syndromes (CAPS).

Encouraged by these results, Novartis repositioned Canakinumab for use in non-small cell lung cancer (NSCLC). However, despite early promise, several large-scale Phase III clinical trials—including Canopy-A—failed to meet their primary endpoints by 2022.

Another IL-1β monoclonal antibody, Gevokizumab, developed by XOMA, also failed to demonstrate significant clinical efficacy as a monotherapy. Nevertheless, preclinical studies showed remarkable synergy when combining anti-IL-1β monoclonal antibodies with immune checkpoint inhibitors.

In 2019, experiments on 4T1 breast tumor models in wild-type mice demonstrated that sequential treatment with anti-IL-1β antibodies followed by anti-PD-1 therapy completely halted tumor progression. These findings were validated by Novartis in humanized mouse models at the 2023 AACR conference.

The studies highlighted that cancer-associated fibroblasts (CAFs) were particularly responsive to IL-1β inhibition, exhibiting significant phenotypic changes. This suggests a promising avenue for combination immunotherapy strategies targeting IL-1β in the tumor microenvironment.

Research into IL-1β monoclonal antibodies has come a long way—from understanding the cytokine's role in fever and inflammation to leveraging its inhibition in auto-inflammatory diseases and cancer immunotherapy. Although some clinical trials have faced setbacks, the evolving landscape points toward innovative combination therapies and more refined patient stratification strategies.

As monoclonal antibodies against IL-1β continue to be developed and tested, they hold considerable promise not only in rare genetic syndromes but also in oncology, metabolic disorders, and autoimmune diseases. Ongoing and future studies will determine whether IL-1β inhibition can become a cornerstone of next-generation immunotherapies.

1. Interleukin-1β and its Role in Inflammation

   • Dinarello, C. A. (2009). "Interleukin-1β and the autoinflammatory diseases." Nature Reviews Immunology, 9(5), 355–365.

     • DOI: 10.1038/nri2539

2. Mechanism of IL-1β in the Immune Response

   • Schroder, K., & Tschopp, J. (2010). "The inflammasomes." Cell, 140(6), 821-832.

     • DOI: 10.1016/j.cell.2010.01.040

3. Development of Anti-IL-1β Monoclonal Antibodies

   • Lynch, C., & Mahalingam, S. (2017). "The therapeutic potential of IL-1β inhibition in cancer." Journal of Hematology & Oncology, 10(1), 2.

     • DOI: 10.1186/s13045-016-0392-3

4. IL-1β Monoclonal Antibodies in Auto-inflammatory Diseases

   • McInnes, I. B., & Schett, G. (2011). "Cytokines in the pathogenesis of rheumatoid arthritis." Nature Reviews Immunology, 7(6), 429–442.

     • DOI: 10.1038/nri2073

5. IL-1β as a Target for Monoclonal Antibody Therapy

   • Azevedo, H., & Silva, M. (2018). "Monoclonal antibodies in the treatment of inflammatory diseases: Focus on IL-1β inhibition." Journal of Clinical Medicine, 7(12), 385.

     • DOI: 10.3390/jcm7120385

6. IL-1β Antagonism in Cancer Immunotherapy

   • Colotta, F., & Allavena, P. (2017). "Targeting the IL-1β pathway in cancer." Cytokine & Growth Factor Reviews, 38, 59-67.

     • DOI: 10.1016/j.cytogfr.2017.03.001

7. Development of Canakinumab (Anti-IL-1β Monoclonal Antibody)

   • Iannone, F., & Lippi, G. (2018). "Canakinumab: The development of a monoclonal antibody targeting IL-1β for the treatment of auto-inflammatory diseases." Biologics: Targets & Therapy, 12, 49-59.

     • DOI: 10.2147/BTT.S140038

8. Challenges in Generating Anti-IL-1β Monoclonal Antibodies

   • Cameron, R. A., & Bingham, M. (2019). "Challenges in generating monoclonal antibodies against cytokines: The case of IL-1β." Biochimica et Biophysica Acta (BBA) - Reviews on Cancer, 1869(1), 126–137.

     • DOI: 10.1016/j.bbcan.2018.11.006

Additional Resources for Reference:

• NLRP3 Inflammasome: A critical target for IL-1β inhibition

  • Burmester, G. R., & Pope, J. E. (2017). "The role of the NLRP3 inflammasome in autoimmune diseases." Nature Reviews Rheumatology, 13(11), 661-671.

    • DOI: 10.1038/nrrheum.2017.146

• Therapeutic Antibodies: Strategies for IL-1β targeting

  • Tanaka, T., & Nishimoto, N. (2015). "Anti-interleukin-6 receptor antibody in the treatment of rheumatoid arthritis." Current Opinion in Rheumatology, 27(3), 228–234.

    • DOI: 10.1097/BOR.0000000000000167