Pooled Protein Immunization for Identification of Cell Surface Antigens in Streptococcus sanguinis 英文参考文献.docVIP
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Pooled Protein Immunization for Identification of Cell Surface Antigens in Streptococcus sanguinis 英文参考文献
PooledProteinImmunizationforIdentificationofCell
SurfaceAntigensinStreptococcussanguinis
XiuchunGe1,ToddKitten1,2,3,CindyL.Munro4,DanielH.Conrad3,PingXu1,2,3*
1PhilipsInstituteofOralandCraniofacialMolecularBiology,VirginiaCommonwealthUniversity,Richmond,Virginia,UnitedStatesofAmerica,2CenterfortheStudyof
Biological Complexity, Virginia Commonwealth University, Richmond, Virginia, United States of America, 3Department of Microbiology and Immunology, Virginia
CommonwealthUniversity,Richmond,Virginia,UnitedStatesofAmerica,4DepartmentofAdultHealthNursing,VirginiaCommonwealthUniversity,Richmond,Virginia,
UnitedStatesofAmerica
Abstract
Background: Available bacterial genomes provide opportunities for screening vaccines by reverse vaccinology. Efficient
identificationofsurfaceantigensisrequiredtoreducetimeandanimalcostinthistechnology.Wedevelopedanapproach
toidentifysurfaceantigensrapidlyinStreptococcussanguinis,acommoninfectiveendocarditiscausativespecies.
Methods and Findings: We applied bioinformatics for antigen prediction and pooled antigens for immunization. Forty-
seven surface-exposed proteins including 28 lipoproteins and 19 cell wall-anchored proteins were chosen based on
computeralgorithmsandcomparativegenomicanalyses.Eightproteinsamongthesecandidatesand2otherproteinswere
pooledtogethertoimmunizerabbits.TheantiserumreactedstronglywitheachproteinandwithS.sanguiniswholecells.
Affinitychromatographywasusedtopurifytheantibodiesto9oftheantigenpoolcomponents.CompetitiveELISAand
FACS results indicated that these 9 proteins were exposed on S. sanguinis cell surfaces. The purified antibodies had
demonstrableopsonicactivity.
Conclusions: The results indicate that immunization with pooled proteins, in combination with affinity purification, and
comprehensiveimmunologicalassaysmayfacilitatecellsurfaceantigenidentificationtocombatinfectiousdiseases.
Citation: Ge X, Kitten T, Munro CL, Conrad DH, Xu P (2010) Pooled Protein Immunization for Identification of Cell Surface Antig
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