A lithographic process is used to form a plurality of target structures (T) on a substrate (W). Each target structure comprises overlaid gratings each having a specific overlay bias. Asymmetry (A) of each grating, measured by scatterometry, includes contributions due to (i) the overlay bias, (ii) an overlay error (OV) in the lithographic process and (iii) bottom grating asymmetry within the overlaid gratings. Asymmetry measurements are obtained for three or more target structures having three or more different values of overlay bias (e.g., −d, 0, +d). Knowing the three different overlay bias values and a theoretical curve relationship between overlay error and asymmetry, overlay error (OV) can be calculated while correcting the effect of bottom grating asymmetry. Bias schemes with three and four different biases are disclosed as examples. Gratings with different directions and biases can be interleaved in a composite target structure.