Fig 1.
(A) Schematic representation of the preparation process for KY-NAb-GDF15, a neutralizing antibody against GDF15. (B-C) ELISA assays evaluating the binding and blocking activities of KY-NAb-GDF15. (B) Both KY-NAb-GDF15 and Ponsegromab exhibited significant binding activity towards GDF15, with EC50 values of 0.05080 μg/mL and 0.1095 μg/mL, respectively. (C) Both KY-NAb-GDF15 and Ponsegromab demonstrated effective blocking activity against the interaction between GDF15 and GFRAL, with IC50 values of 0.2525 μg/mL and 0.5037 μg/mL for KY-NAb-GDF15 and Ponsegromab, respectively. (D) KY-NAb-GDF15 inhibits GDF15-stimulated GFRAL signaling in the HEK293 SRE-luc2-cRET-GFRAL reporter system with higher efficiency than Ponsegnomab. (E) With 0.1 μg/mL antibody concentration, KY-NAb-GDF15 (EC50 = 16.2 ng/mL) neutralized more GDF15 than Ponsegnomab (EC50 = 11.80 ng/mL).
Fig 2.
Analysis of the specific binding of KY-NAb-GDF15.
(A) ELISA assay evaluating the interaction between KY-NAb-GDF15 and GDF family proteins. (B-E) Flow cytometry analysis assessing the binding affinity of KY-NAb-GDF15 towards peripheral blood mononuclear cells (B), umbilical vein endothelial cells (C), red blood cells (D), and granulocytes (E).
Table 1.
Stability assessment of KY-NAb-GDF15 antibody using SEC-HPLC and ELISA techniques following various treatments.
The stability testing of KY-NAb-GDF15 was performed under different acceleration conditions, including 5 freeze-thaw cycles, low pH treatment, high-temperature treatment for 1 week and 2 weeks, as well as stability analysis in plasma. Before and after treatment, KY-NAb-GDF15 was analyzed using SEC-HPLC and ELISA to detect changes in integrity and binding function.
Fig 3.
In vivo pharmacokinetic assessment of KY-NAb-GDF15.
BALB/c mice received intravenous injections of the antibody KY-NAb-GDF15, and serum antibody levels were quantified using ELISA. The half-life (t1/2) of KY-NAb-GDF15 was determined to be approximately 10.14 days at a concentration of 3 mg/kg.
Fig 4.
Neutralizing effect of KY-NAb-GDF15 on GDF15-mediated weight loss.
(A) KY-NAb-GDF15 mitigates GDF15-induced reduction in body weight. BALB/c mice received GDF15-mFc injections on day 0, followed by administration of KY-NAb-GDF15 or Ponsegromab on day 2. The percentage change in body weight was calculated relative to day 0. (B-C) KY-NAb-GDF15 attenuates tumor-associated weight loss. Body weight changes in HT1080 xenograft mice (B) and LS513 xenograft mice (C) following intraperitoneal (IP) injection of KY-NAb-GDF15 or Ponsegromab at doses of 1 mg/kg and 10 mg/kg. The percentage change in body weight was calculated relative to day 0. (D) KY-NAb-GDF15 alleviates chemotherapy-induced weight loss. GDF15 humanized mice (B-hGDF15) or WT C57BL/6 mice were treated with cisplatin and KY-NAb-GDF15 or hIgG. The percentage change in body weight was calculated relative to day 0.