A　facile　strategy　based　on　the　synergistic　effect　of　molecular　imprinting　and　boronate　affinity　was　proposed　for　glycoprotein　imprinting.　Polydopamine　(PDA)-coated　boronic　acid-functionalized　molecularly　imprinted　silica　nanoparticles　(MIPs)　were　prepared　by　the　＂thiol-ene＂　click　reaction　using　SiO2　as　the　core,　3-acrylamidophenyl　boronic　acid　(AAPBA)　as　the　functional　monomer,　and　horseradish　peroxidase　(HRP)　as　the　glycoprotein　template.　A　well　defined　core-shell　structure　of　MIPs　was　obtained　after　self-polymerization　of　dopamine　(DA)　on　the　surface　of　HRP-immobilized　silica　nanoparticles　(NPs).　The　polymerization　conditions　and　adsorption　behavior　were　investigated　in　detail　in　order　to　obtain　the　highest　selectivity　and　binding　capacity.　Under　the　optimized　conditions,　the　HRP-MIPs　showed　higher　binding　affinity　towards　HRP　than　non-imprinted　nanoparticles　(NIPs),　and　the　corresponding　adsorption　capacity　(Q)　and　imprinted　factor　(α)　reached　0.58　μmol　g-1　and　2.6,　respectively.　The　specificity　for　HRP　recognition　was　evaluated　with　a　competitive　experiment,　and　the　results　indicated　that　the　HRP-MIPs　had　higher　selectivity　for　the　template.　The　good　features　of　the　HRP-MIPs　facilitated　selective　isolation　and　enrichment　of　trace　HRP　from　human　serum.　In　addition,　the　stability　and　regeneration　were　also　investigated,　which　indicated　that　the　HRP-MIPs　had　excellent　reusability.　©　2015　The　Royal　Society　of　Chemistry.