Flow injection analysis of intracellular beta-galactosidase in Escherichia coli cultivations, using an on-line system including cell disruption, debris separation and immunochemical quantification.
A continuous integrated process for on-line quantification of intracellular components has been developed. By applying the concept of expanded micro-beds in a flow injection system it was possible to first perform on-line cell disintegration followed by an on-line binding assay for quantification of a reporter protein (beta-galactosidase) from the cell interior. The disintegration process involved the use of an expanded bed with immobilised lysozyme followed by ultrasonic treatment in a flow-through cell. The cell debris does not interfere in the binding assay as it is carried out in an expanded bed. The time for an assay cycle is at present approx. 35 min. This integrated system can be used for quantification of proteins down to at least 10(-7) mol/L.
Tocaj
A
A
Department of Biotechnology, Lund University, Sweden.
Nandakumar
M P
MP
Holst
O
O
Mattiasson
B
B
D002138:Calibration; D004926:Escherichia coli / Q000201:enzymology*; D017022:Flow Injection Analysis / Q000379:methods*; D007150:Immunohistochemistry; D014465:Ultrasonics; D001616:beta-Galactosidase / Q000032:analysis*
D001616:beta-Galactosidase