B/B0 Ratio at Standard Curve Endpoint Out of Range

Symptom

The B/B0 ratio (bound/maximum bound) at the lowest or highest standard concentration is either >95% or <5%, indicating the standard curve dynamic range is not properly positioned.

1. 1 Standard concentration range does not match assay dynamic range
2. 2 Initial standard concentration too high or too low for the antibody affinity
3. 3 Improper dilution series calculation for competitive format

1. 1 Increase or decrease the starting standard amount by one dilution factor
2. 2 Shift the entire standard curve to reposition within optimal dynamic range
3. 3 Verify standard stock concentration and recalculate dilution series

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