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We demonstrate that ionic self-assembled multilayers (ISAMs) adsorbed on long period gratings (LPGs) function effectively as biosensors using biotin–streptavidin as a demonstration bioconjugate pair. Biotin, streptavidin, and anti-streptavidin were deposited onto the ISAM-coated LPG sequentially, each inducing a measurable resonant wavelength shift of the LPG. Control experiments verified the specificity of the biosensor system. Furthermore, we demonstrate that ISAM coated turnaround point LPGs could serve as highly sensitive biosensors that exhibit a change in transmitted intensity rather than resonant wavelength. Experimental measurements confirm that ISAM-coated LPGs provide an attractive platform for building efficient and high-performance optical sensors.