Shanghai Novamab Biopharmaceuticals Co., Ltd.

TSLP, a cytokine derived from epithelial cells at barrier surfaces, has been extensively studied as a pivotal regulator of type 2 and non-type-2 immune responses, implicated in inflammatory airway diseases. Although mAb therapy targeting TSLP has shown significant therapeutic effects, there is still no approved TSLP-targeting inhaled formulation.

Leveraging the distinct advantages of nanobodies (Nbs) in maintaining efficacy during storage and administration, our objective is to develop a novel inhalable TSLP Nb for effectively treating airway inflammation.

Four TSLP-immunized Nb libraries were screened to generate specific and functional TSLP Nbs. LQ043, a bivalent Nb comprising 2 humanized Nb13 units, was constructed. The efficacy, pharmacokinetics, and safety of inhaled LQ043 were evaluated in mice and nonhuman primates, followed by a randomized, double-blind phase I clinical study. The clinical trial was registered with the Chinese Clinical Trial Registry (CTR20230092).

LQ043 exhibited high affinity and specificity for human TSLP, coupled with a favorable inhibitory effect on TSLP/TSLP receptor interaction. Furthermore, inhaled LQ043 significantly attenuated ovalbumin-induced allergic inflammation in mice and demonstrated excellent safety, pharmacokinetic characteristics, and tissue distribution in nonhuman primate models, with high concentrations observed in the respiratory system. Notably, phase I clinical trial results demonstrated dose-proportional pharmacokinetics, safety, and good tolerability for LQ043. In addition, LQ043 can be produced on a large scale at low cost in yeast cells, thereby substantially alleviating the economic burden associated with inhaled biopharmaceuticals.

These preclinical and clinical studies robustly establish the safety and efficacy of inhaled LQ043, underscoring its potential as a pioneering inhalation therapy for airway inflammation.

Cytokines are promising in cancer immunotherapy, but their pleiotropic effects limit specificity and clinical utility. Through binding to IL-18Rα and IL-18Rβ, interleukin-18 (IL-18) stimulates innate lymphocytes and effector T cells for antitumor immunity. However, clinical trials of recombinant IL-18 have been hampered by IL-18 binding protein (IL-18BP), a secreted high-affinity decoy receptor. Here, we developed decoy-resistant bispecific nanobodies that maintain IL-18 signaling potential. Based on agonistic nanobodies targeting IL-18Rα and IL-18Rβ, bispecific nanobody A4B2-mdFc effectively enhanced CD8⁺ T cell responses with distinct transcriptomic profiles. Systemic delivery of A4B2-mdFc boosted CD8⁺ T cell infiltration and activation, demonstrating dose-dependent antitumor efficacy in cell-line-derived xenograft and patient-derived xenograft models. Interestingly, PDCD1 and CTLA4 expressions were drastically increased on CD8⁺ T cells when treated with A4B2-mdFc. Injection of A4B2-mdFc significantly improved the antitumor efficacy of immune checkpoint inhibitors (ICIs) targeting both PD-1 and CTLA-4. Our findings demonstrated that nanobody-based bispecific IL-18 mimetics elicited superior antitumor activity via CD8⁺ T cell activation and IL-18BP resistance, providing the potential application of cytokine-targeting bispecific nanobody monotherapy or in combination with ICIs for cancer immunotherapy.

Adeno-associated virus serotype 8 (AAV8) is a widely used gene therapy vector with the characteristics of high transduction efficiency and tissue specificity. Reliable detection of AAV8 is crucial for assessing therapeutic efficacy and tracking its in vivo distribution. Herein, we develop a nanobody-based electrochemiluminescence (ECL) imaging immunosensor based on integrating advanced functional materials with spatially resolved signal output for sensitive and visual detection of AAV8. This immunosensor employs a tetraphenylethylene-derived covalent organic framework (TC-COF) as the ECL emitter with the advantages of excellent chemical stability, structural rigidity, and aggregation-induced luminescence. Moreover, the gold-rhodium core-shell bimetallic nanoparticles (Au@Rh) are introduced as the efficient coreaction accelerators to promote the formation of triethylamine (TEA) radicals. Nanobodies serve as the high-affinity and low-steric-hindrance recognition elements with the advantages of enhanced epitope accessibility and stable surface immobilization. Both the Au@Rh and nanobodies contribute to good specificity and high sensitivity. Notably, this ECL immunosensor enables direct visualization of AAV8 binding events, and the pixel-based signal quantification ensures robust calibration and high reproducibility. This ECL immunosensor can achieve a wide linear detection range from 1 × 10⁸ to 5 × 10¹¹ vg/mL, with the limit of detection (LOD) reaching as low as 1 × 10^7.15 vg/mL. To the best of our knowledge, this work reports the first application of AAV8-specific nanobodies in an ECL biosensing platform. This study demonstrates a significant analytical advancement by integrating nanobody technology, COF-based luminescent materials, and catalytic nanoparticles into a robust and broadly applicable sensing platform for virus detection.