TY - JOUR
T1 - On-chip Escherichia coli culture, purification, and detection of expressed proteins
AU - Kim, Moonil
AU - Lee, So Young
AU - Choi, Hyunju
AU - Shin, Yong Beom
AU - Jung, Sun Ok
AU - Kim, Min Gon
AU - Chung, Bong Hyun
PY - 2006/10
Y1 - 2006/10
N2 - 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.
AB - 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.
KW - Glutathione S-transferase-fused green fluorescent protein (GST-GFP)
KW - Microwell chip
KW - On-chip purification
KW - Surface plasmon resonance (SPR) imaging system
UR - http://www.scopus.com/inward/record.url?scp=33749264992&partnerID=8YFLogxK
U2 - 10.1007/s00249-006-0072-8
DO - 10.1007/s00249-006-0072-8
M3 - Article
C2 - 16724194
AN - SCOPUS:33749264992
SN - 0175-7571
VL - 35
SP - 655
EP - 662
JO - European Biophysics Journal
JF - European Biophysics Journal
IS - 8
ER -