Polyphosphazene　microspheres　(PZS)　multispectral　encoded　by　fluorescence　and　surface　enhanced　Raman　spectra　(SERS)　were　prepared　and　used　as　solid　phase　carrier　of　capture　antibody　for　antibody　detection.　A　variety　of　fluorescent　encoded　polyphosphazene　microspheres　(FPZS)　incorporating　single　and　multiple　fluorescent　molecules　were　prepared.　Multiple　SERS　encoded　polyphosphazene　microspheres　(SPZS)　were　obtained　by　modification　of　different　Raman　reporters　on　the　surface　of　PZS@Ag,　which　were　fabricated　with　rough　morphology　through　in　situ　deposition　of　Ag　nanoparticles.　Fluorescence　and　SERS　spectral　combined　encoded　polyphosphazene　microspheres　(FSPZS)　were　prepared　by　encoding　of　fluorescent　molecules　and　SERS　reporters.　The　optical　signals　in　the　two　independent　detection　channels　were　separable　and　ensured　the　accuracy　for　the　encoding　method.　Immunoassays　of　goat　IgG,　rabbit　IgG　and　mouse　IgG　were　performed　by　the　as-prepared　FPZS,　SPZS　and　FSPZS　as　solid　phase　carrier　of　capture　antibody　and　fluorescent　labeled　second　antibody,　using　sandwich　immunoassay　method.　The　immunoassay　results　showed　that　the　multiple-spectra　encoded　microspheres　have　stable　and　distinguishable　encoding　signals,　large　encoding　volume　and　multiplexed　analytical　capability　in　sandwich　immunoassays.　©　2021,　The　Author(s),　under　exclusive　licence　to　Springer-Verlag　GmbH　Germany,　part　of　Springer　Nature.