This study presents a method to enhance proton conductivity in hydrogen-bonded organic frameworks (HOFs) through vitrification. By rapidly quenching the kinetically stable HOF-SXU-8 to a glassy state (HOF-g), proton conductivity was significantly improved without the need for humidity. The vitrified HOF-g exhibited a proton conductivity of 5.62 × 10⁻² S cm⁻¹ at 100 °C, a dramatic increase from the 1.31 × 10⁻⁷ S cm⁻¹ of the original HOF-SXU-8. The glassy HOF-g also showed excellent long-term stability, maintaining high conductivity even at 30 °C. Molecular dynamics (MD) simulations and X-ray total scattering experiments revealed that the HOF-g system consists of three types of clusters: 1,5-Naphthalenedisulfonic acid (1,5-NSA) anion clusters, N,N-dimethylformamide (DMF) molecule clusters, and H⁺-H₂O clusters. The H⁺ ions play a crucial role in bridging these clusters, contributing to the high proton conductivity. The study demonstrates that vitrification can effectively eliminate grain boundary effects in HOFs, enabling efficient proton conduction and advancing energy conversion and storage technologies. The findings provide valuable insights for optimizing HOFs for proton conduction applications.This study presents a method to enhance proton conductivity in hydrogen-bonded organic frameworks (HOFs) through vitrification. By rapidly quenching the kinetically stable HOF-SXU-8 to a glassy state (HOF-g), proton conductivity was significantly improved without the need for humidity. The vitrified HOF-g exhibited a proton conductivity of 5.62 × 10⁻² S cm⁻¹ at 100 °C, a dramatic increase from the 1.31 × 10⁻⁷ S cm⁻¹ of the original HOF-SXU-8. The glassy HOF-g also showed excellent long-term stability, maintaining high conductivity even at 30 °C. Molecular dynamics (MD) simulations and X-ray total scattering experiments revealed that the HOF-g system consists of three types of clusters: 1,5-Naphthalenedisulfonic acid (1,5-NSA) anion clusters, N,N-dimethylformamide (DMF) molecule clusters, and H⁺-H₂O clusters. The H⁺ ions play a crucial role in bridging these clusters, contributing to the high proton conductivity. The study demonstrates that vitrification can effectively eliminate grain boundary effects in HOFs, enabling efficient proton conduction and advancing energy conversion and storage technologies. The findings provide valuable insights for optimizing HOFs for proton conduction applications.