On-chip Escherichia coli culture, purification, and detection of expressed proteins.
In a recent study, we reported the results of a rapid high-throughput expression analysis of the affinity-tagged proteins present in total cell lysates, using a surface plasmon resonance (SPR) imaging protein chip system. In this paper, we describe a novel method, which is able to sequentially carry out a recombinant Escherichia coli culture, as well as the detection and purification of the expressed proteins on a single microwell chip, fabricated on a two-dimensional thin gold film. Following the induction of the protein on the microwell chip, the E. coli cells were lysed on the chip via the addition of lysozymes, and the expressed glutathione S-transferase-fused green fluorescent protein (GST-GFP) was then purified on the chip via affinity interaction with the glutathionylated gold surface of the chip. Finally, the expressed protein was directly detected using the surface plasmon resonance (SPR) imaging system. This system saves a substantial amount of time, experimental resources, and labor, by allowing for the complicated and labor-intensive procedures inherent to the production of recombinant proteins to be conducted on a single microwell chip, simply and economically.
Kim
Moonil
M
BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, 115, Yuseong, Daejeon, South Korea.
Lee
So-Young
SY
Choi
Hyunju
H
Shin
Yong-Beom
YB
Jung
Sun Ok
SO
Kim
Min-Gon
MG
Chung
Bong Hyun
BH
D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D049452:Green Fluorescent Proteins / Q000235:genetics; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D020349:Surface Plasmon Resonance
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005982:Glutathione Transferase