蛋白质印迹模板的制备及在尿液蛋白测定中的应用
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摘要
以牛血清蛋白(BSA)为分子模板,甲基丙烯酸(MAA)为功能单体,乙二醇二甲基丙烯酸酯(EDGMA)为交联剂,偶氮二异丁腈(AIBN)为引发剂,制备了BSA分子印迹模板,研究了该模板对BSA的吸附性能,并以其作为净化材料去除人尿中的干扰物质,测定了人尿中的蛋白质。结果表明:蛋白质模板吸附和洗脱的最佳pH值分别为5.3和5.6,最佳吸附时间为2h,最大吸附容量为72mg/g。三种人尿样品中蛋白质的含量分别为35.4、18.9、24.2mg/L,回收率为92.4%~103.7%。该方法可用于人尿中蛋白质含量的测定。
The protein imprinted polymer is synthesized with bovine serum albumin(BSA)as template,methacrylic acid(MAA)as functional monomer,ethylene glycol dimethacrylate(EGDMA)as a crosslinker,and azobisisobutyronitrile as initiator.Afterwards,the protein imprinted material was used as purification materials for determination of protein in human urine.The results show that the best absorption and clean pH value is 5.3and 5.6,respectively,and the adsorption capacity is 72 mg/L.Moreover,the concentration of protein in three human urine samples were determined to be 35.4,18.9and 24.2 mg/L,with the recovery between 92.4% and 103.7%.The method was suitable for the determination of protein in human urine.
The protein imprinted polymer is synthesized with bovine serum albumin(BSA)as template,methacrylic acid(MAA)as functional monomer,ethylene glycol dimethacrylate(EGDMA)as a crosslinker,and azobisisobutyronitrile as initiator.Afterwards,the protein imprinted material was used as purification materials for determination of protein in human urine.The results show that the best absorption and clean pH value is 5.3and 5.6,respectively,and the adsorption capacity is 72 mg/L.Moreover,the concentration of protein in three human urine samples were determined to be 35.4,18.9and 24.2 mg/L,with the recovery between 92.4% and 103.7%.The method was suitable for the determination of protein in human urine.
引文
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[16]WANG Shou-xing(王守兴),WANG Feng(王峰),HUANG Wei(黄薇).Journal of Analytical Science(分析科学学报)[J],2007,23(4):464.
[2]LI Ling-ling(李玲玲).Journal of Chongqing Industry&Trade Polytech(重庆工贸职业技术学院学报)[J],2011,21(1):47.
[3]PEI Cao-yu(裴朝玉),FANG Guo-bo(方国波).Journal of Zhoukou Normal University(周口师范学院学报)[J],2010,23(3A):78.
[4]WU Wen-xiang(吴文镶).Journal of Langfang Teachers University(廊坊师范学院学报)[J],2014,20(4):11.
[5]Lowry O H,Rosebrough N J,Farr A L.Biol Chem[J],1951,193:265.
[6]Bradford M M.Anal Biochem[J],1976,72(1):248.
[7]Flores R.Anal Biochem[J],1978,88(2):605.
[8]Guo Z X,Shen H X.Analyst[J],1999,124(7):1093.
[9]DONG Xue-zhi(董学芝),DENG Cui-zhen(邓翠贞),XIN Jian-hao(信建豪).Chin J Pharm Anal(药物分析杂志)[J],2008,28(6):928.
[10]ZHANG Zheng-qi(张正奇),CHEN Yong-xiang(陈永湘),ZHANG Yu-cong(张郁葱).Journal of Analytical Science(分析科学学报)[J],2001,17(3):214.
[11]CUI Feng-ling(崔凤灵),WANG Jun-li(王俊丽),CUI Yan-rui(崔延瑞).Spectroscopy and Spectral Analysis(光谱学与光谱分析)[J],2008,28(2):38 4.
[12]QIN Meng(秦蒙),LIANG Shu-cai(梁淑彩),CHEN Jia-qi(陈珈琪).Journal of Analytical Science(分析科学学报)[J],2014,30(2):227.
[13]Li Z P,Fan S S,Zhang L N.Anal Sci[J],2004,20(9):1327.
[14]XIONG Yang-hui(熊阳辉),HU Yong-gang(胡涌刚),PANG Zhi-tao(庞志涛).Physical Testing and Chemical Analysis Part B:Chemical Analysis(理化检验-化学分册)[J],2008,44(3):231.
[15]ZHANG Qiang-zhai(张强斋),YUN Yan-ru(贠嫣茹),CUI Feng-ling(崔凤灵).Journal of Henan Normal University(河南师范大学学报)[J],2011,39(3):82.
[16]WANG Shou-xing(王守兴),WANG Feng(王峰),HUANG Wei(黄薇).Journal of Analytical Science(分析科学学报)[J],2007,23(4):464.