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BT20 cells yielded high EC50 values for the two antibodies, which also correlated with the lowest specific maximal lysis observed (compare Tables 3 and 4).
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Lysis was expressed as a percentage of the lysis achieved with 1% Triton X-100 after subtraction of spontaneous lysis observed in the absence of test Ab.
However, when the CWI of ER and ISO12 were evaluated experimentally with an enzymatic cell wall lysis assay (lyticase) no significant differences in maximal lysis rate were observed (Additional file 1).
Cytotoxicity was calculated using the following formula: percent cytotoxicity = (experimental lysis − spontaneous lysis) ÷ (maximal lysis − spontaneous lysis) × 100.
Haemolysis is presented as a percentage of maximal lysis by H2O.
Percentage of specific lysis was calculated as follows: (experimental cpm − basal cpm)/(maximal cpm − basal cpm) × 100, with maximal lysis determined in the presence of 5% triton and basal lysis in the absence of antibody and effectors.
Results were expressed as % specific lysis determined as [ cytotoxic release - Min)/(Max - Min)] × 100, where the spontaneous lysis corresponds to the minimal release (Min), and the lysis provoked by addition of hydrochloric acid corresponds to the maximal lysis (Max).
Optimal labelling time had previously been determined by choosing the labelling time with the highest maximal lysis:background radioactivity ratio.
C50 values (relative tonicity at half maximal lysis) were calculated by fitting the data to a 4-parameter logistic sigmoidal curve.
The maximal value observed is 0.35.
Convex hulls were drawn to delineate the area of maximal variation observed in the samples.
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