Protein constituency of unbound fraction of bovine serum applied to anion exchange columns at different pHs and molarities with or without ethylenediamine tetraacetic acid. Application to preparation of bovine IgG1.

Ethylenediamine tetraacetic acid disodium salt (EDTA) was found to be an important addition to anion exchange buffers in terms of the profile and amounts of eluted bovine serum proteins. Hydrogen ion concentration and buffer composition were important when different types of anion exchanger (DEAE 52 cellulose and QAE Sephadex A 50) were used for separation of bovine serum proteins as functional groups of the different anion exchangers did not behave similarly and were therefore not interchangeable. These findings applied to the purification of bovine serum IgG1 amounting to 15 to 25% of the total IgG1 was accomplished using QAE Sephadex A 50 anion exchange chromatography. This was followed by absorption of the IgG1 fraction with Staphylococcus aureus containing protein A to remove minor IgG2 contaminants and gel filtration to exclude traces of the third component of complement (C3).