A Blind Spot in Alignment: Quantifying Biosecurity Risks in Large Language Models

Large Language Models (LLMs) are accelerating biological research, yet this same capability poses a critical biosecurity threat: models that assist in protein engineering can equally be prompted to design functional toxins, potentially lowering the barrier to biological misuse. Current safety evaluations, however, operate in natural language and cannot determine whether a model-generated amino acid sequence is biological gibberish or a credible threat. To address this evaluation blind spot, we introduce SPIKE-Bench, coupling 631 curated toxin-design prompts across seven functional categories with the SPIKE funnel, a three-stage protocol that filters output through compliance, biological plausibility, and predicted toxicity, producing stage-level diagnostics and an aggregate function-aware metric: the Functional Harmfulness Rate (FHR). An audit of 32 LLMs reveals that most models freely comply with toxin-design requests; FHR is driven primarily by biological generation capability rather than safety alignment, reaching 50.7\%; and Refusal Rate fails to predict functional risk. As a first step toward mitigation, we provide BioSafe-Guard, a domain-specialized classifier that substantially reduces predicted functional risk while preserving benign utility. We release SPIKE-Bench and BioSafe-Guard at \url{https://anonymous.4open.science/r/SPIKE-Bench} to support more rigorous biosecurity evaluation of LLMs.