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1、上海交通大學(xué)碩士學(xué)位論文分別為IgG1和IgG2a,通過體內(nèi)誘生腹水法獲得純化后效價為1:64000的抗軟骨藻酸單克隆抗體。3.膠體金及軟骨藻酸金標(biāo)抗體的制備采用檸檬酸三鈉還原氯金酸法,成功確立了20nm膠體金的制備條件,并利用制備好的膠體金,對前述制備的抗軟骨藻酸單克隆抗體進(jìn)行了膠體金標(biāo)記條件的摸索,成功確立了單克隆抗體的膠體金標(biāo)記條件:最適pH是9.0,最適標(biāo)記穩(wěn)定量是9.72μg/mL膠體金,并制備金標(biāo)抗體。4.軟骨藻酸膠體金免疫層析試紙條的制備對軟骨藻酸膠體金免疫層析試紙條的各項指標(biāo)進(jìn)行優(yōu)化,并確立了最佳條件。最終選擇SB06玻璃纖維素膜涂布4×稀釋度的金標(biāo)抗體構(gòu)成膠體金結(jié)合墊;選
2、擇Millipore180NC膜作為層析膜,羊抗鼠IgG(1mg/mL)和1μL包被抗原DA-BSA(0.867mg/mL)噴涂在NC膜上,分別作為質(zhì)控線和檢測線;選用SB08玻璃纖維素膜作為樣品墊;SX18吸水紙作為吸收墊;將膠體金結(jié)合墊、NC膜、樣品墊和吸收墊組裝成軟骨藻酸膠體金免疫層析試紙條5.軟骨藻酸膠體金免疫層析試紙條的應(yīng)用對膠體金免疫層析試紙條的檢測靈敏度、特異性和穩(wěn)定性進(jìn)行測試,結(jié)果表明,試紙條檢測時間不超過15min,檢測靈敏度達(dá)20ng/mL(對于貝類食品相當(dāng)于8μg/g貝肉),可以滿足國際規(guī)定的20μg/g的安全限值。特異性強、重復(fù)性好,4℃下可保存6個月。該方法靈敏度
3、高、簡單快速,無需特殊儀器設(shè)備,適用于廣大基層單位的現(xiàn)場和批量檢測,有著良好的商業(yè)開發(fā)應(yīng)用前景。關(guān)鍵詞:記憶缺失性貝毒,軟骨藻酸,單克隆抗體,膠體金,免疫層析第II頁萬方數(shù)據(jù)上海交通大學(xué)碩士學(xué)位論文DEVELOPMENTOFCOLLOIDALGOLDIMMUNOCHROMATOGRAPHICSTRIPFORRAPIDDETECTIONOFAMNESICSHELLFISHPOISONINGTOXINABSTRACTAmnesicShellfishPoisoning(ASP)isakindoftoxinthatproducedbytoxicredtidediatoms.Themaincompo
4、undinvolvedinthistoxicityisdomoicacid(DA),anexcitatoryneurotransmitteraminoacidofthekainicgroupthathadbeenisolatedfromamarinesponge.Domoicacidcanbebio-concentratedbyshellfishthroughthetransmissionoffoodchain,andcanalsobedischargedintomarinewaterdirectlybydiatoms.Peopleandothermarinemammalswhotakei
5、nthecontaminatedseafoodscouldsufferfromcentralnervoussystemdysfunction,notablylossofshort-termmemory,evencomaanddeath.Colloidalgoldimmunochromatographicassay(GICA)isanewimmunochromatographictechniqueinwhichacellulosemembraneisusedasthecarrier,andacolloidalgold-labeledantigenorantibodyisusedasthetr
6、acer.Itissimple,rapidandcheap,andtheresultisclearandeasytoanalyzewithoutanyequipment/facilitiesandskilledtechnicians.Sinceitisverysuitableforspot,rapiddetectionandbatchdetectiononsite,theGICAtechniquehasbecomeoneofthefastestgrowingimmunologicaldetectionmethodsandiswidelyusedforrapiddetectioninmari
7、netoxinsanalysis.Inthisstudy,thecolloidalgoldimmunochromatographicstripforrapiddetectionofdomoicacid(DA)wasdevelopedbyusingtheprincipleofmonoclonalantibodyandimmunochromatographicassay.Theresultswere第III頁萬方數(shù)據(jù)上海交通