A method is provided for separating a protein from one or more other
proteins using hydroxyapatite chromatography in which the protein does
not bind to hydroxyapatite but the other protein(s) does. In some
embodiments, a second protein affixed to a solid support has been used
previously to purify the protein by affinity chromatography, and small
amounts of the second protein are introduced in the sample during this
process. The protein being purified can comprise at least one constant
antibody immunoglobulin domain. The second protein can bind to proteins
comprising such a domain.