纤维素磁性微球的制备及其生物活化研究  

STUDY ON PREPARATION AND BIOACTIVATION OF NOVEL IMMUNOMAGNETIC MICROSPHERES

作　　者：孙丹[1] 曹宇[2] 白钢[2,3] 张奇[3] 

机构地区：[1]南开大学学报编辑部,天津300071 [2]南开大学生命科学学院,天津300071 [3]南开大学药学院,天津300071

出　　处：《离子交换与吸附》2009年第1期17-24,共8页Ion Exchange and Adsorption

基　　金：天津市应用基础及前沿技术研究计划(08JCYBJC04400)

摘　　要：通过基因重组技术将增强型绿色荧光蛋白(EGFP)基因插入含有纤维素结合域(CBD)标签的载体pET-35b(+),使EGFP与CBD进行融合表达,表达的融合蛋白分子量约为53kDa,该蛋白既具有与纤维素结合的功能,又能够作为示踪信号。此外,通过反向悬浮包埋技术制备纤维素磁性微球(MCMS),并对其性能进行了考察,其平均粒径为(158.7±14.7)μm,磁含量为(3.83±0.29)%,磁响应时间为(66±2.9)s,磁漏损为(0.139±0.06)‰,蛋白的偶联量和漏损量分别为(12.27±0.82)mg/mL和(77±7.35)ng/mL。经由EGFP的示踪信号观察制备的MCMS与CBD具有良好的结合能力,吸附后微球表面具有明显的绿色荧光信号。本研究建立的CBD-MCMS将为后继的MCMS的生物活化和随后的分离、纯化等工作奠定基础。The gene of enhanced green fluorescence protein （EGFP） was inserted into pET-35b（＋） including cellulose binding domain （CBD） by gene engineering method in this study. And then the fusion protein CBD-EGFP was expressed in E. coli BL21. The molecular mass was 53kDa, approximately. The fusion protein owned two functions： binding cellulose and being signal of tracking. At the same time, we prepared magnetic cellulose microspheres （MCMS） and characterized them. The mean diameter was （158.7±14.7）pm, the magnetic content was （3.83±0.29）%, the response time of magnetic is （66±2.9）s and the magnetic leakage was （0.139±0.06）‰. The protein contents of binding and leakage were （12.27±0.82）mg/mL and （77±7.35）ng/mL, respectively. Using tracking signal from EGFP, we observed the clear fluorescence signal at MCMS surface. It showed that CBD could bind MCMS well and firmly. The CBD-MCMS system, which was established by us, could provide one base for biological activation and purification of protein, and so on.

关 键 词：纤维素磁性微球 纤维素结合域 荧光蛋白 生物偶联 

分 类 号：TQ425.2[化学工程]