Separation and concentration of bacteria with immobilized antibody fragments

• Published in: Journal of applied bacteriology Vol. 78; no. 4; p. 359
• Main Authors: Molloy, P, Brydon, L, Porter, A J, Harris, W J
• Format: Journal Article
• Language: English
• Published: England 01.04.1995
• Subjects: Antibodies, Bacterial - biosynthesis; Antibodies, Bacterial - immunology; Antibody Affinity; Base Sequence; Chromatography, Affinity; Cloning, Molecular; DNA, Bacterial; Escherichia coli; Food Microbiology; Immunoglobulin Fab Fragments - biosynthesis; Immunoglobulin Fab Fragments - immunology; Immunoglobulin Fragments - biosynthesis; Immunoglobulin Fragments - immunology; Molecular Sequence Data; Pseudomonas aeruginosa - immunology; Pseudomonas aeruginosa - isolation & purification
• ISSN: 0021-8847
• DOI: 10.1111/j.1365-2672.1995.tb03418.x

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.