Oligo(dT)亲和层析介质的载量比较和机制分析  

Binding capacity and mechanism analysis of four Oligo(dT)affinity chromatography resins

作　　者：谭远志 张鹏程[1] 孙艳娜 张其磊[1] 姚善泾[1] 林东强[1] TAN Yuanzhi;ZHANG Pengcheng;SUN Yanna;ZHANG Qilei;YAO Shanjing;LIN Dongqiang(Key Laboratory of Biomass Chemical Engineering of Ministry of Education,College of Chemical and Biological Engineering,Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学生物质化工教育部重点实验室,浙江大学化学工程与生物工程学院,浙江杭州310058

出　　处：《高校化学工程学报》2024年第2期243-252,共10页Journal of Chemical Engineering of Chinese Universities

基　　金：国家重点研发计划(2021YFC2103401,2021YFE0113300);国家自然科学基金(22078286)。

摘　　要：针对Oligo(d T)亲和层析介质的吸附性能,以poly(A)为模型分子,考察了4种Oligo(d T)亲和层析介质的静态吸附平衡、吸附动力学和动态结合载量(DBC),探讨了载量影响相关机制。结果表明,4种介质的合适吸附条件均为0.6 mol·L-1Na Cl、p H=6~7;Monomix d T20静态吸附容量最大,且poly(A)能扩散至介质微球深层孔内,而Poros Oligo(d T)25、Praesto Jetted (d T)25和Nano Gel d T20等3种介质中poly(A)均主要为表层吸附、静态吸附容量稍低;对于DBC,Nano Gel d T20和Monomix d T20的10%穿透的DBC较高,而Poros Oligo (d T)25和Praesto Jetted (d T)25相对略低。经分析,影响载量的主要因素包含基质种类、微球孔径、配基密度、间隔臂和配基长度等。对于基质种类,聚苯乙烯基质可能孔道结构较为特别。对于微球孔径,应针对不同大小的m RNA分子定制不同孔径的微球,以平衡传质阻力与可及吸附表面积之间的矛盾,从而增大DBC。In order to study binding performance of four Oligo(dT)affinity chromatography resins,static adsorption equilibria,adsorption kinetics and dynamic binding capacities(DBC)were investigated with poly(A)as the model target.The results show that the optimal static adsorption capacities of the four resins were all obtained at 0.6 mol·L^(-1) NaCl and pH 6-7.The Monomix dT20 resin showed the highest static adsorption capacity and poly(A)molecule could diffuse into the pores of resin beads.For Poros Oligo(dT)25,Praesto Jetted(dT)25 and NanoGel dT20,poly(A)molecule mainly bounded to the surface of resin beads,which resulted in a slightly lower static adsorption capacity.The DBC results show that the DBC10%of NanoGel dT20 and Monomix dT20 were relatively high,while those of Poros Oligo(dT)25 and Praesto Jetted(dT)25 were slightly lower.The main factors affecting the binding capacity were found as matrix type,microsphere pore size,ligand density,spacer arm structure and ligand length.Polystyrene-based resins might have special pore structure.For mRNA molecules of varying sizes,resin beads with suitable pore sizes should be tailored to balance mass transfer resistance and effective adsorption surface area to obtain optimal DBC results.

关 键 词：亲和层析 m RNA 寡脱氧胸腺苷酸 载量 生物分离 

分 类 号：TQ028.8[化学工程]