USE OF BIOLUMINESCENCE FOR THE EVALUATION OF AFFINITY CONSTANTS FOR BACTERIAL CELL-ANTIBODY INTERACTIONS

For live bacterial cell assays, it is important to choose antibodies that provide high assay sensitivities. To accomplish this, the affinity constants of antibodies raised against target bacteria need to be determined and compared. Bacterial surfaces provide a vast array of antigenic determinants such as polysaccharides, proteins, and glycolipids. The affinity constants of antibodies raised against these small, extracted, molecular components can be easily derived, however it is uncommon to determine that against the bacterial cell as a whole. As a further complication, when antibodies are raised against bacteria, the cells are heat-treated and thus may have a different affinity constant when reacted to live cells. If the assay is used to detect live bacteria, it is pertinent to choose antibodies with high affinity constants that have been tested against bacteria with intact physiology…