Antigenic sites on a protein have been predicted by the measurement of hydrophilicity values. We have compared this approach to the use of atomic temp factors to predict the antigenic sites on insulin. These predictions were based upon computer assisted analysis of X-ray crystallography data. The results demonstrate that the hydrophilic sites A6, B19, B23, B24, and B26 are not on the surface of the molecule and therefore are not potential antigenic determinants. Thus, hydrophilicity values alone are not sufficient to predict antigenic sites. In contrast, atomic temp factors were more predictive of antigenicity. These findings may explain why anti-insulin antibodies are less likely to develop against certain hydrophilic sites.