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1、維普資訊http://www.cqvip.com第18卷第1期江蘇大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)V01.18No.12008年1月JournalofJiangsuUniversity(MedicineEdition)Jan.2008Ⅱ型膠原蛋白抗血清的制備和檢測(cè)姜旭淦,朱敏娟,葛彥文,吳亮,陳盛霞,許化溪(江蘇大學(xué)醫(yī)學(xué)技術(shù)學(xué)院,江蘇鎮(zhèn)江212013)[摘要]目的:探討Ⅱ型膠原蛋白(collagenproteintypeⅡ,CⅡ)抗體產(chǎn)生規(guī)律,制備CⅡ抗血清。方法:采用純化的雞和豬CⅡ免疫新西蘭白兔,每隔10d收集血清,以改良聞接ELISA法檢測(cè)抗CⅡ抗體效價(jià)。結(jié)果:雞CⅡ免疫新西蘭白兔后,
2、第10天抗體效價(jià)為1:3200,第20天升至1:204800;加強(qiáng)免疫后,第30天和第40天抗體效價(jià)分別達(dá)到1:409600和1:819200。豬CⅡ免疫新西蘭白兔后,第10天血清中未能檢測(cè)出抗體,第20天血清抗體效價(jià)為1:3200;加強(qiáng)免疫后,第30天和第40天抗體效價(jià)分別為1:51200和1:102400。結(jié)論:采用純化的雞和豬CI1免疫新西蘭白兔后,經(jīng)一定潛伏期血清中出現(xiàn)抗體,可以獲得高效價(jià)抗CⅡ抗體血清??贵w產(chǎn)生情況符合機(jī)體初次及再次免疫應(yīng)答的有關(guān)規(guī)律,但血清抗體產(chǎn)生的潛伏期和效價(jià)與不同種屬來(lái)源的膠原蛋白有關(guān)。[關(guān)鍵詞]雞;豬;Ⅱ型膠原蛋白;抗Ⅱ型膠原蛋白抗體;酶聯(lián)免疫
3、吸附實(shí)驗(yàn)[中圖分類號(hào)]R446.6[文獻(xiàn)標(biāo)志碼]A[文章編號(hào)]1671—7783(2008)O1—0034—03PreparationanddetectionofantiseraforcollagenproteintypeⅡJIANGXu-gan,ZHUMin-juan,GEYah-wen,刪Liang,CHENSheng-xia,XUHua-xi(SchoolofMedicalTechnology,JiangsuUniversity,ZhenjiangJiangsu212013,China)[Abstract]Objective:Toinvestigatetheregular
4、patternsofantibodyproductionandpreparetheantise-rumoncollagenproteintypeII(CII).Methods:TheNewZealandwhiterabbitswereimmunizedwithpu-rifledchickenandpigCIIandthentherabbitserawerecollectedevery10days.TheimprovedindirectELISAmethodwasadoptedtodetectthetitersofanti.CIIantibodies.Results:Whenth
5、erabbitswereim.munizedwithpurifiedchickenCII。theantibodytitersinserawere1:3200at10thdayand1:204800at20thday.Afterboostertheantibodytitersincreasedto1:409600at30thdayand1:819200at40thday.WhentherabbitswereimmunizedwithpurifiedpigCII,theantibodiescouldnotbedetectedat10thday.Butat20thdaytheanti
6、bodytiterwas1:3200.Afterboostertheantibodytiterswerethenincreasedto1:51200at30thdayand1:102400at40thday.Conclusion:Theanti.CIIantibodiescanemergeinser-umafterlatencyandtheantiserumwithhightitercanbeobtainedwhentherabbitswereimmunizedwithpu-rifledchickenandpigCII.Theantibodyproductionwasconsi
7、stentwithrelevantregularityofprimaryandsecondaryimmuneresponses.Thelatentphasesandtitersofanti.CIIantibodiesinserawereconcernedwithcollagenproteinsfromdifferentgenus.[Keywords]chicken;pig;collagenproteintypeII;anti-collagenproteintypeIIantibody;enz