Separation of bispecific antibodies and bispecific antibody production by-products using hydroxyapatite chromatography

摘要

The present disclosure is directed to a method that includes the use of hydroxyapatite chromatography to separate the bispecific antibody from a solution that also contains one or more byproducts characteristic of bispecific antibody production. A byproduct unique to the production of bispecific antibodies (bispecific antibody specific byproducts, `` BASB '') includes fragments of the bispecific antibody and heavier molecular weight variants of the antibody, Fragments and / or the variants contain an Fc domain but do not show affinity for two different epitopes and / or antigens exhibited by the desired bispecific antibody. Accordingly, the methods of the present disclosure include separation of the bispecific antibody from one or more of the bispecific antibody BASBs. The hydroxyapatite chromatography method of the present disclosure may be used alone or to achieve any level of purity of the bispecific antibody required for, for example, therapeutic and / or diagnostic applications. It may be further combined with standard purification steps and unit operations known in the art.