抗伏馬菌素B-,1-單鏈抗體的制備.pdf

1、福建農(nóng)林大學(xué)碩士學(xué)位論文抗伏馬菌素B單鏈抗體的制備姓名：劉曉雷申請學(xué)位級別：碩士專業(yè)：微生物學(xué)指導(dǎo)教師：汪世華20100401福建農(nóng)林大學(xué)碩士學(xué)位論文PreparationofSingleChainAntibodyvariableFragmentagainstFumonisinBIAbstractFumonisin，akindofmycotoxinderivedfromFusariummoniliforme，canpollutecrop

2、sandtheirproductswidelyIthasdiversetoxicitythatthreatensthehealthofhumanbeingsandanimalsThisstudyintendedtoestablishamethod，whichWasbasedonrecombinantsinglechainantibodyagainstfumonisinBI，todetectthecontentsoffumonisinBl

3、infoodandfeedquicklyandefficientlyFBlisamicromoleculeandhapten，SOitneedstocouplewithamacromoleculeproteinandtheconjugatewillbecomeacompleteantigen’Inthisresearch，fumonisinBl(FB1)Waspurchased，andglutaraldehyde(GA)one—step

4、methodwasusedtoprepareFBIBSAconjugateandGAtwo—stepmethodtoprepareFBI’KLHconjugateFBlBSAwasusedasimmunizingantigenandFB1KLHWasusedfordetection，andthebothconjugateswereprovedbythefollowingtiterassayresultsofantiserumBalb／c

5、micewereimmunized晰tllFBI—BSAconjugate，andthetiterofmiceantiserumwasassayedbyELISAafterboosterwhichachievedtheexpectationThen，theimmunizedmousewaskilledanditsspleenwasseparatedTotalRNAWasextractedfromthespleenandthefirsts

6、trandofcDNAWasobtainedbyreversetranscriptionAndthegenes％and圪，fromcDNAtemplatewereusedtoconstructgenescFvSubsequently，phageantibodylibrarywasestablishedafterthegenescFvWasclonedintophageplasmidpCANTAB5EScFvpanningfromthel

7、ibrarybyphagedisplaytechniqueswascarriedoutThesolubleexpressionandidentificationofscFvwereperformedFinally，weobtainedonestrainofphageantibodybeingabletoproducescFvwithhighaffinityKeyWords：fumonisin；singlechainantibody；ph