Development of an oxalate-affinity chromatographic column to isolate oxalate-binding proteins

[摘要] Calciumoxalate(CaOx)isthemajorcrystallinecomponentinkidneystones.Manypreviousstudiesonstonemodulationhavefocusedmainlytocalcium-bindingproteins,whereasoxalate-bindingproteinsremainunder-investigatedduetoalackofasimplemethodtopurifyoxalate-bindingproteins.Therefore,wehavedevelopedanaffinitychromatographiccolumntoisolateorpurifyoxalate-bindingproteins.Theoxalate-affinitycolumndevelopedcontainedEAH-Sepharose4Bbeadsconjugatedwithoxalicacidbycarbodiimidereactionusing1-ethyl-3-(3-dimethylaminopropyl)-carbodiimidehydrochloride(EDC)asanactivator.Thecouplingefficacyofcoupledbeads(oxalicacid-EAHSepharose4B)wasdeterminedbyacompetitiveninhydrinassaytoquantifytheremainingaminogrouponEAH-Sepharose4B.Inaddition,theoxalate-affinitycolumnshowedhighspecificbindingtoaknownoxalate-bindingprotein(p62)butnottoanirrelevantprotein(carbonicanhydrase).Inconclusion,wehaveestablishedtheoxalate-affinitychromatographiccolumn,whichshowedthehighefficiencytoisolateoxalate-bindingproteins.Thisnovelchromatographiccolumnwillbeveryusefulforfuturestoneresearch,particularlyintheareaofstonemodulationbyoxalate-bindingproteins...

[学科分类] 分析化学