富勒醇-fitc磷光标记固体基质室温磷光法测定痕量碱性磷酸酶

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1、2011年第1期漳州师范学院学报(自然科学版)No．1．2011匀i(总71期)JournalofZhangzhouNormalUniversity(Nat．Sci．)GeneralNO．71文章编~：1008．7826(2011)01．0063．07富勒醇一FITC磷光标记固体基质室温磷光法测定痕量碱性磷酸酶林少琴(福建教育学院理科研修部，福建福州350001)摘要：开发了新的富勒醇、异硫氰酸荧光素和N，N．二甲基苯胺(F．oi．(FITC)n。DMA)标记试剂．基于F．ol的活性一OH与FITC游离的-COOH反应生成含有多个FITC

2、分子的复合物(F—oi．(FITC)n。DMA)，增加WGA—AP—WGA—F—olFITC-DMA(WGA和AP分别为麦胚凝集素与碱性磷酸酶)生物靶的发光分子数而进一步提高SSRTP(固体基质室温磷光法)的灵敏度．用F—o1．(FITC)n—DMA标记WGA方法的检出限为O．15agspot。’(F-o1)与0．097agspot((FITC)，比F—ol—DMA、FITC-DEA单发光分子标记WGA方法的检出F~-(O．20agspot(F．o1)．0．22agspot(FITC)低．用于血清试样中AP含量的测定结果与酶联免疫法相吻合

3、．关键词：碱性磷酸酶；富勒醇一畀硫氰酸荧光素；麦胚凝集索；亲和吸附固体基质室温磷光法中图分类号：0629．8文献标识码：ASolidSubstrateRoomTemperaturePhosphorimetryfortheDeterminationofTraceAlkalinePhosphataseUsingFullerol—fluoresceinIs0thi0cyanateasLabellingReagentLINShao-qin(DepartmentofScienceTrainingandResearching，FujianEducat

4、ionCollege，Fuzhou，Fujian350001，China)Abstract：Anewph0sph0rescentlabellingreagentconsistingoffulleroi，fiuoresceinisothiocyanateandN，N—dimethylanilinef

5、—ol一(FITC)n—DMA)wasdeveloped．Themodeofactionisbasedonthereactivityoftheactive··OHgroupinF··olwiththe--COOHgroupofF

6、TCtofor

7、manF·-ol·-(FITC)n--DMAcomplexcontainingseveralFITCmolecules．F—ol一(FITC1n—DMAincreasedthenumberofluminescentmoleculesinthebiologicaltargetofWGA-AP-WGA-F—ol一(FITC)n—DMA(WGAandAParewheatgermagglutininandalkalinephosphatase，respectively)whichimprovedthesensitivityusingsolidsu

8、bstrateroomtemperaturephosphorimetry(SSRTP)detection．TheproposedmethodprovidedhighsensitivityandstrongspecificityforWGA-AP．Thelimitofdetection(LD)wasO．15agAPspot～forF—olandO．097agAPspot—forFITCinF-ol一(FITC)n-DMA，whichwaslowerthanthemethodusingsingleluminescentmoleculesofF

9、-oI-DMAandFITC-DMAtolabelWGAr0．20agAPspot～forF-ol-DMAand0．22agAPspot～forFITC—DMA)．ResultsforthedeterminationofAPinhumanserumwereingoodagreementwiththoseobtainedbyenzyme·linkedimmunosorbentassay．ThemechanismofF—oi一(FITC)n-DMAlabellingofWGAwasdiscussed．Keywords：Alkalinephos

10、phatase；Fullerol-fluoresceinis0thiocyanate；Wheatgermagglutinin；Afinityadsorptionsolidsubstratero