摘要
构建了一种基于渗透脱水模式的自动进样微流控结晶芯片.该芯片通过真空预脱气将包含蛋白质和结晶剂的液滴自动分配至结晶微腔阵列中,然后利用集成的一排包含不同浓度盐溶液的透析管道,通过渗透脱水方式经一层聚二甲基硅氧烷(PDMS)膜实现液滴的逐渐浓缩,使之趋于过饱和状态,进而形成结晶.此芯片可一次筛选较宽范围的过饱和状态,实现蛋白质结晶的快速优化.利用模式蛋白溶菌酶的结晶实验验证了该芯片的性能。
A self-dispensing and osmosis-based microfluidic crystallization device was reported. This device automatically dispenses droplets containing protein and precipitant into an array of crystallization chambers by pre-degassing, and then gradually concentrates these droplets at different rates by osmotic dewatering through a polydimethylsiboxane（PDMS） membrane using a row of integrated dialysis channels filled with different con- centration of salt solution. This concentration process drives the protein/precipitant mixture into supersatura- ted, thus tending toward the crystallization state. This device allows for screening a wide range of supersatura- tion in one trial for rapid optimization of protein crystallization. The feasibility of this crystallization device is demonstrated using the model proteins of lysozyme.
出处
《高等学校化学学报》
SCIE
EI
CAS
CSCD
北大核心
2012年第10期2178-2183,共6页
Chemical Journal of Chinese Universities
基金
国家“九七三”计划项目(批准号:2011CB707505,2012CB933303)
国家自然科学基金(批准号:60906055,61006086和30900315)
中国科学院科研装备研制项目(批准号:YZ201143)
上海市自然科学基金(批准号:10ZR1436200)资助
关键词
自动进样
高通量筛选
渗透脱水
蛋白质结晶
Self-dispensing
High-throughout screening
Osmotic dewatering
Protein crystallization
