血清蛋白预处理方案对双向电泳效果的影响  

Influence of different pre-treatments for serum on the effects of two-dimensional electrophoresis

作　　者：李天旺[1] 黄志祥[1] 郑奔容[1] 邓伟明[1] 王敏[1] 古洁若[1] 

机构地区：[1]中山大学附属第三医院风湿内科,广东广州510630

出　　处：《中国医学工程》2011年第1期1-4,8,共5页China Medical Engineering

基　　金：广东省科技计划项目(2008B030301039)

摘　　要：目的比较不同的血清蛋白预处理方案对双向凝胶电泳(2-DE)分辨率与重复性的影响。方法收集健康志愿者血清样本3例,分别以原始血清及经Aurum Serum Protein Mini Kit(ASPMK)、ProteoMiner Pro-tein Enrichment Kit(PMPEK)、Multiple Affinity Removal System(MARS)处理后的血清样本进行2-DE,所得2-DE图谱经PDQuest软件分析,比较上述4种方法所获得的凝胶图谱的整体蛋白质点数及其变异系数(CV)、组内凝胶匹配点数。同时,通过划分白蛋白、转铁蛋白、纤维蛋白原、IgA与α1-抗胰蛋白酶富集区域(Ⅰ区)及IgG分布区域(Ⅱ区),细化比较其中蛋白点数。结果 3名健康志愿者的原始血清及经ASPMK、PMPEK、MARS处理的血清样本2-DE图谱中蛋白质点数分别为589±83、611±45、798±26及810±20个,CV分别为41.1%、28.6%、23.5%及22.9%,组内匹配蛋白质点数分别为424±50、523±34、713±18及734±19个。在Ⅰ区和Ⅱ区,经PMPEK及MARS处理的血清样本蛋白质点数均较经ASPMK处理的样本获原始血清多。结论预处理可提高血清样本2-DE分辨率及重复性;以PMPEK及MARS处理血清效果优于AS-PMK。【Objective】To compare the resolution and reproducibility of different pre-treatments for serum in two-dimensional electrophoresis（2-DE）.【Methods】Three serum samples were collected from different healthy volunteers and each were pre-treated using Aurum Serum Protein Mini Kit（ASPMK）,ProteoMiner Protein Enrichment Kit（PMPEK） and Multiple Affinity Removal System（MARS） respectively.Then both the raw and treated serum samples were subjected to 2-DE.The 2-DE gels were analyzed by PDQuest softwar,and the total number of protein spots,its coefficient of variation in sets and the number of matched spots were compared among different pre-treatment methods.Additionally,number of spots in regions which albumin,transferring,fibrinogen,IgA,α1-antitrypsin enriched（Region Ⅰ） and IgG distributed（Region Ⅱ） were analyzed respectively.【Results】The total spot numbers in the gels of raw serum,serum which have been treated with ASPMK,PMPEK and MARS were 589±83,611±45,798±26 and 810±20,and CV were 41.1%,28.6%,23.5% and 22.9% respectively.The matched spots numbers were 424±50,523±34,713±18 and 734±19 in the above group.In both of Region Ⅰand Region Ⅱ,the numbers of protein spot in gels run with samples treated with PMPEK and MARS were significantly greater than those with raw serum and sample treated with ASPMK.【Conclusion】The pre-treatment might improve the resolution and reproducibility of 2-DE.PMPEK and MARS are better than ASPMK in pre-treatment of serum.

关 键 词：血清 双向凝胶电泳 蛋白质组学 

分 类 号：R457.2[医药卫生—治疗学]