Method for assessment of functional affinity of antibodies for live bacteria.
A rapid and convenient method for assessment of functional affinity of antibodies against live bacteria is described. When a combination of immunomagnetic separation (IMS) with bioluminescent or fluorescent genetic labelling of the cells was employed, the method showed good correlation with plate count. However, the use of reporter bacteria allowed results to be obtained within 1 h compared with days using conventional methods. Due to its lower detection limit, the bioluminescent assay performed better than the fluorescent assay. Antibody affinities for Escherichia coli O157:H7 and Salmonella enteritidis were examined at different environmental conditions such as pH 3-7, temperature 4-25 degrees C, and sodium chloride concentrations 0-5% and compared with sensitivities of ELISA.
Brovko
Lubov
L
Canadian Research Institute for Food Safety, University of Guelph, 43 McGilvray str. Guelph, Ontario, Canada N1G 2W2. lbrovko@uoguelph.ca
Young
Denise
D
Griffiths
Mansel W
MW
D000907:Antibodies, Bacterial / Q000276:immunology*; D000915:Antibody Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D019453:Escherichia coli O157 / Q000276:immunology* / Q000302:isolation & purification; D018189:Immunomagnetic Separation; D008163:Luminescent Measurements; D008856:Microscopy, Fluorescence; D012044:Regression Analysis; D012477:Salmonella enteritidis / Q000276:immunology* / Q000302:isolation & purification
D000907:Antibodies, Bacterial