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Cited by 7 publications
(8 citation statements)
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“…Maximum fluorescence (Q max ) was defined as the unliganded fluorophore and observed fluorescence (Q obs ) was defined as the liganded fluorophore. Percentage decrease from maximum [(1 À (Q obs / Q max ))  100%] was calculated from corrected data 52 and further normalized by defining the final saturated titration fluorescence as 100% quenching. Binding affinity was calculated by using the Langmuir binding equation, Y ¼ (B max  X)/(K d þ X) and the GraphPad Prizm 3.0 software, where B max is maximal binding, and X is total DNA concentration in molarity.…”
Section: Fluorescence Measurementsmentioning
confidence: 99%
“…Maximum fluorescence (Q max ) was defined as the unliganded fluorophore and observed fluorescence (Q obs ) was defined as the liganded fluorophore. Percentage decrease from maximum [(1 À (Q obs / Q max ))  100%] was calculated from corrected data 52 and further normalized by defining the final saturated titration fluorescence as 100% quenching. Binding affinity was calculated by using the Langmuir binding equation, Y ¼ (B max  X)/(K d þ X) and the GraphPad Prizm 3.0 software, where B max is maximal binding, and X is total DNA concentration in molarity.…”
Section: Fluorescence Measurementsmentioning
confidence: 99%
“…Phage clones recovered from library selections are then individually validated for target binding; yet, this is usually a time-consuming step that can strongly benefit from new methods to rapidly prioritize hits. Some examples of increasing throughput include a fluorescence-based method for rapid ranking of recovered clones (Landon et al 2004a), and a computational approach for the graphical display of ELISA data to depict affinity and specificity rankings (Kelly et al 2006a). If desired, additional mutagenesis can be introduced into the recovered phage DNA to further improve properties such as binding affinity Friedman et al 2008).…”
Section: Selections Against Purified Targets Intact Cells and Ex VImentioning
confidence: 99%
“…The maximum fluorescence (Q max ) is defined as fluorescence at 348 nm of the free GDH and observed fluorescence (Q obs ) is defined as the fluorescence after addition of titrant. The decrease in fluorescence emission (1-(Q obs /Q max )) × 100 %) with different concentration of azo-NAD + is fit to a Langmuir binding model[32] (Equation 3) to obtain the binding affinities. Y=Bmax[normalA]([A]+Kd) where B max and [A] are the maximum specific binding and ligand concentration respectively.…”
Section: Methodsmentioning
confidence: 99%