Bovine　serum　albumin　imprinted　polyethylene　glycol　600　(PEG600)　grafted　Calcium　alginate　(CaA)　hydrogel　microspheres　were　prepared　and　characterized.　The　adsorption　and　recognition　properties　of　PEG600　grafted　calcium　alginate　(CaA-g-PEG600)　microspheres　were　evaluated　and　the　results　showed　that　the　crosslinking　structure　of　CaA-g-PEG600　microspheres　exerted　an　obvious　effect　on　the　adsorption　capacity　and　imprinting　properties　for　bovine　serum　albumin.　The　adsorption　isotherms　and　recognition　properties　indicated　that　the　imprinted　modified　microspheres　had　excellent　rebinding　affinity　toward　target　proteins　and　the　imprinting　efficiency　varied　according　to　PEG600　grafting　degree.　The　adsorption　capacity　and　the　imprinting　factor　were　5.5　mg　g−1　and　3.6,　respectively.　Adsorption　kinetics　of　CaA-g-PEG600　microspheres　in　accordance　with　the　molecular　weight　between　crosslinks　(Mc)　was　investigated　and　the　structural　influence　on　protein　selective　rebinding　was　discussed.　Furthermore,　the　binary　solution　separation　performance　of　CaA-g-PEG600　microspheres　with　different　Mc　was　investigated　by　selective　binding　bovine　serum　albumin　from　protein　mixture　solution.　©　2017,　Springer　Science+Business　Media　Dordrecht.