Separation and concentration of bacteria with immobilized antibody fragments.
New methods to quantitatively remove bacteria from food and water samples are required to meet modern safety standards. The recent development of techniques to make Fab/Fv/scFv fragments in bacteria has provided the opportunity to exploit antibodies as specialized chemicals for affinity removal technologies. Single-chain fragments against Pseudomonas aeruginosa have been expressed in Escherichia coli, purified via a fused poly-histidine tail and immobilized upon polystyrene beads. The resulting immunoaffinity columns have been shown to effectively remove greater than 90% of an applied 10 million bacteria after a single passage through the column. Column material in the absence of single-chain retained less than 10% of the bacteria. Pseudomonas were also removed from milk, mixed bacterial cultures and when present at low cell densities.
Molloy
P
P
Department of Molecular and Cell Biology, University of Aberdeen, Marischal College, UK.
Brydon
L
L
Porter
A J
AJ
Harris
W J
WJ
D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000276:immunology*; D000915:Antibody Affinity; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli; D005516:Food Microbiology; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000276:immunology*; D008969:Molecular Sequence Data; D011550:Pseudomonas aeruginosa / Q000276:immunology / Q000302:isolation & purification*
D000907:Antibodies, Bacterial; D004269:DNA, Bacterial; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments