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《A Structure-Based Database of Antibody Variable Domain Diversity》由会员上传分享，免费在线阅读，更多相关内容在行业资料-天天文库。
1、doi:10.1016/j.jmb.2005.02.063J.Mol.Biol.(2005)348,699–709AStructure-BasedDatabaseofAntibodyVariableDomainDiversity1*,ChristianWiesmann21ChristopherJ.Bond,JamesC.MarstersJrand2*SachdevS.Sidhu1DepartmentofMedicinalThediversityofnaturalantibodiesislimitedbythegeneticmechanismsChemistry,GenentechI
2、nc.thatengenderdiversityandthefunctionalrequirementsofantigen1DNAWay,SouthSanbinding.Usinganinvitro-evolvedautonomousheavychainvariableFrancisco,CA94080USAdomain(VHH-RIG),wehaveinvestigatedthelimitsofstructurally-2tolerateddiversityinthethreecomplementarity-determiningregionsandDepartmentofPro
3、teinafourthloopwithinthethirdframeworkregion.WedeterminedtheX-rayEngineering,GenentechInc.crystalstructureoftheVHH-RIGdomainat1.9A˚resolutionanduseditto1DNAWay,SouthSanguidethedesignofphage-displayedlibrariesencompassingthefourloops.Francisco,CA94080USAThelibrariesweresubjectedtoselectionsfors
4、tructuralstability,andadatabaseofstructurally-toleratedsequenceswascompiledfromthesequencesofapproximately1000uniqueclones.Theresultsrevealthatallfourloopsaccommodatesignificantlygreaterdiversitythanisobservedinnature.Thus,itappearsthatmostsequencebiasesinthenaturalimmunerepertoirearisefromfact
5、orsotherthanstructuralconstraintsand,consequently,itshouldbepossibletoenhancethefunctionsofantibodiessignificantlythroughinvitroevolution.q2005ElsevierLtd.Allrightsreserved.Keywords:phagedisplay;proteinengineering;combinatorialmutagenesis;*Correspondingauthorsantibody;variabledomainIntroduction
6、variabledomainsisgeneratedbydistinctgenetic9–13mechanisms.Atthelevelofthegermline,Thevertebrateimmunesystemcanevolveanti-diversityisencodedinthecollectionofVgenes;bodiescapableofrecognizingessentiallyanyattheleveloftheB-cell,variationisengenderedbymacromoleculewithhighaffinityandspecificity.reco
7、mbinationoftheV,DandJsegments.Recom-Analysesofnaturalantibodysequences,togetherbinationgeneratesvariabilityinaminoacidcontent,withstructuralstudies,havebeeninstrumentalinandforCDR3,inlooplengthaswell.Finally,1–3revealinghowantibodieswor
