The affinity will influence the slope of the curve. Beware: in the detection of specific antibodies it’s not that easy to reach because of the fact that two properties influencing the ELISA signal: the antibody concentration and the affinity. You have to make sure that your assay has a dilution linearity (one sample in different dilution has to give the same final result: i.e. This can be done by the definition of a positive pool which has 100 arbitrary units. The standards can be set easily either by an international standard available via NIBSC or by a "self made standard". If you add two quality controls (high and low level) you can assure that the measurement is correct.
If quantitative, use as said before a calibration with a set of at least 5 standards and a respective calibration curve.
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