Open AccessCrystallizing proteins on the basis of their precipitation diagram determined using a microfluidic formulator
Author(s) -
Sommer Morten O. A.,
Larsen Sine
Publication year - 2005
Publication title -
journal of synchrotron radiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.172
H-Index - 99
ISSN - 1600-5775
DOI - 10.1107/s0909049505002621
Subject(s) - crystallization , protein crystallization , microfluidics , phase (matter) , precipitation , protein precipitation , protein kinase a , chemistry , materials science , crystallography , computer science , biological system , chromatography , biochemistry , nanotechnology , kinase , biology , physics , organic chemistry , meteorology , analyte
Crystallization of proteins from a purified protein solution remains a bottleneck in the structure determination pipeline. In this paper the crystallization problem is addressed using a microfluidic device capable of determining detailed protein precipitation diagrams using less than 10 µL of protein sample. Based on the experimentally determined protein phase behavior, a crystallization screen can be designed to accommodate the physical chemistry of the particular protein target. Such a tailor‐made crystallization screen has a high probability of yielding crystallization hits. The approach is applied to two different proteins: the calcium pump (SERCA), an eukaryotic integral membrane protein, and UMP kinase, a prokaryotic soluble kinase. Protein phase behavior is mapped for both proteins and tailor‐made crystallization screens are designed for the two proteins resulting in about 50% crystallization probability per experiment. This illustrates the power of using microfluidic devices for detailed characterization of protein phase behavior prior to crystallization trials.