Abstract In the event of a public health crisis, highly effective and sustainable antimicrobial materials and equipment will be urgently needed. Here, the preparation is reported by electrospinning of broad‐spectrum antibacterial nanofibers embedded in a photoactive hydrogen‐bonded organic framework (HOF) of rod‐like nanocrystals ≈60 nm in length. The resulting HOF@PVDF‐HFP nanofibers maintain excellent tensile and breathability characteristics while shielding HOF nanocrystals against acid and alkali corrosion. A series of nanofibers embedded with different amounts and types of HOF nanocrystals are prepared to optimize their efficiency of singlet oxygen ( 1 O 2 ) generation. The 0.5 wt.% HOF‐101‐F@PVDF‐HFP nanofibers exhibit the most efficient 1 O 2 generation that is enhanced by a factor of almost 2 when compared with the HOF‐101‐F microcrystalline powder. The HOF@PVDF‐HFP nanofibers are demonstrated to highly effectively kill pathogens, including viruses, bacteria, and fungi in 30 min under ambient light conditions.