ResearchpaperAnenzyme-linkedimmunosorbentassay(ELISA)forquantificationofhumancollectin11(CL-11,CL-K1)L.Selmana,M.L.Henriksena,J.Brandta,Y.Palarasaha,A.Watersb,P.L.Bealesb,U.Holmskovc,T.J.D.Jørgensend,C.Nielsene,K.Skjodta,S.Hansena,⁎aDepartmentofCancerandInflammationResearch,InstituteofMolecularMedicine,UniversityofSouthernDenmark,Winslowparken21-1,DK-5000Odense,DenmarkbMolecularMedicineUnit,InstituteofChildHealth,UniversityCollegeLondon,30GuilfordStreet,LondonWC1N1EH,UKcInstituteofMolecularMedicine,UniversityofSouthernDenmark,Winslowparken25-3,DK-5000Odense,Odense,DenmarkdDepartmentofBiochemistryandMolecularBiology,UniversityofSouthernDenmark,Campusvej55,DK-5230OdenseM,DenmarkeDepartmentofClinicalImmunology,OdenseUniversityHospital,Sdr.Boulevard29,DK-5000OdenseC,DenmarkarticleinfoabstractArticlehistory:Received14September2011Receivedinrevisedform13October2011Accepted13October2011Availableonline20October2011Collectin11(CL-11),alsoreferredtoascollectinkidney1(CL-K1),isapatternrecognitionmoleculethatbelongstothecollectingroupofproteinsinvolvedininnateimmunity.Itinter-actswithglycoconjugatesonpathogensurfacesandhasbeenfoundincomplexwithmannose-bindinglectin-associatedserineprotease1(MASP-1)and/orMASP-3incirculation.MutationintheCL-11genewasrecentlyassociatedwiththedevelopmentalsyndrome3MC.Inthepresentstudy,weestablishedandthoroughlyvalidatedasandwichenzyme-linkedimmu-nosorbentassay(ELISA)basedontwodifferentmonoclonalantibodies.Theassayishighlysensitive,specificandshowsexcellentquantitativecharacteristicssuchasreproducibility,di-lutionlinearityandrecovery(97.7–104%).Theworkingrangeis0.15–34ng/ml.TheCL-11con-centrationintwoCL-11-deficientindividualsaffectedbythe3MCsyndromewasdeterminedtobebelow2.1ng/ml.WemeasuredthemeanserumCL-11concentrationto284ng/mlin100Danishblooddonors,witha95%confidenceintervalof269–299ng/ml.Therewasnosig-nificantdifferenceintheCL-11concentrationmeasuredinmatchedserumandplasmasam-ples.Storageofsamplesandrepeatedfreezingandthawingtoacertainex...
