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Dynamic and on‐site analysis of serum from human blood is crucial, however, state‐of‐the‐art blood‐assay methods can only collect single or discrete data of physiological analytes; thus, the online reports of the dynamic fluctuation of key analytes remains a great challenge. Here, we propose a novel separation‐sensing membrane by constructing a heterogeneous‐nanostructured architecture, wherein a surface nanoporous layer continuously extracts serum, while the biosensing nanochannels underneath dynamically recognise biotargets, thereby achieving a continuous testing of vital clinical indices as blood is drawn. By precisely controlling the pore structure and nanoshape of biosensing crystals, this membrane achieved accurate and online glucose and lactate monitoring in patients with a variety of medical conditions within 1 min, which is one order of magnitude faster than state‐of‐the‐art techniques. Moreover, various kinds of bio‐recognisers can be introduced into this membrane to accurately detect glutamate, transaminase, and cancer biomarkers. [ABSTRACT FROM AUTHOR]