大分子印迹交联聚乙烯基吡咯烷酮微球的制备与特性研究

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1、摘要以N．乙烯基吡咯烷酮(NVP)为单体，羟乙基纤维素(HEC)为分散剂，N，N．亚甲基双丙烯酰胺(bisAM)为交联剂，过氧化苯甲酰／二甲基苯胺(BPO／DMA)为引发剂，牛血清蛋白(BSA)为模板，采用低温悬浮聚合方法，制备了平均粒径为2009m的牛血清蛋白大分子印迹交联聚乙烯基吡咯烷酮微球(BSA-IMs)。讨论了影响BSA．LMs粒径大小、分布和形态的各种因素：水油比、分散剂的种类及加入量、搅拌速度、引发剂的用量及加入方式、反应温度等。BSA．IMs的重结合行为测试结果表明，对牛血清蛋白有较

2、好的特异选择性能，吸附平衡时的印迹效率(IE)可达到3．25。反应温度、BSA中加入盐的浓度、BSA溶液pH值、凝胶微球中模板的洗脱方式对BSA—IMs的吸附量和印迹效率都有不同程度的影响。反应温度会影响微球的形态进而问接影响吸附性能，随着反应温度的升高印迹效率稍有下降；BSA溶液中盐浓度达到某一值时，BSA．1Ms的吸附量出现了极大值，而印迹效率呈现出逐渐下降的趋势；BSA溶液的pH值越低，BSA—IMs的吸附量也越大，但印迹效率逐渐降低；模板的洗脱方式对BSA．IMs的吸附量也有很大影响，用含1

3、．0％SDS的10％乙酸(VⅣ)水溶液洗脱液对BSA．IMs进行洗脱，BSA．IMs体现出了较高的吸附量和印迹效率。关键词：大分子印迹；分子印迹交联聚乙烯基吡咯烷酮微球；低温悬浮聚合；牛血清蛋白；重结合；平衡吸附量；印迹效率ABSTRACTUsingN—vinylpyrrolidone(NVP)asthefunctionalmonomer,N，N'-methylene．bisacrylamide(BisAM)ascrosslinker,benzoylperoxide／amine(BPO／DMA)as

4、theredoxinitiatorsystemandbovineserumalbumin(BSA)astemplate，viasuspensionpolymerization，wepreparedprotein-imprintedcrosslinkedpolyvinyl．pyrrolidone(PVP)hydrogelmicrospheresandwiththediameterofthemicrospheresbeing200pm．Theeffectsoftheratiobetweenthecont

5、inuousphaseandthedispersephase，thestiringspeed，thekindandquantityofthestabilizers，thepolymerizationtemperatureonthesize，sizedistributionandmorphologyofthecrosslinkedPVPmicrospheresintheprocessofpolymerizationwereinvestigated．Theadsorptionandrecognition

6、propertiesofthemicrosphereswerestudied，aswellastheimprintingmechanismwasdiscussed．TheselectivitytestofimprintedmicrospheresshowedthattheimprintedmicrospheresexhibitedgoodrecognitionforBSA．TheadsorptioncapacityofBSA．imprintedmicrosphereswas3．25timesmore

7、thanthatofnon—imprintedmicrospheres．Itwasconsideredthatthef．o加ationofmultiplehydrogenbondsandcomplementaryshapebetweentheimprintingcavitiesandthetemplateproteinswerethetwofactorsthatledtotheimprintingeffect．ThepHofBSAsolution，theionicstrength，theremovi

8、ngmethods，theinitialBSAconcentrationandthepolymerizationtemperaturealldeeplyinfluencedtheadsorptioncapacityandtheimprintingefficiency(IE)oftheBSA．IMs．AddingNa2HP04toBSAsolution，bothoftheBSA．IMsandNon．IMs’equilibriumabsorptioncapacitiesa