![]() ![]() If you are aiming to purify a pharmaceutical product under GMP, I am always afraid of being the first to present new technology to the FDA. It looks like interesting stuff, but I am not sure what the advantages are for your application. I have heard this terminology used often when talking about ion exchange chromatography, I am not sure it is accepted scientific nomenclature. What I mean is : a buffer system where the buffering ion is positively charged (such as Tris) as compared to an Anionic buffer such as carbonate, phosphate or acetate. That is what a forum like this is all about. Sorry if I got side-tracked from your original question, but hopefully someone will learn from my mistakes. There is a reason why you are supposed to use a cationic buffer with anion exchange resins!! When the resin got close to capacity – the citrate had a stronger affinity and displaced the protein. We later determined that during loading, both citrate and antibody were binding to the Q resin. This was lost into the large volume of flow through waste. This worked OK at small scale with a light load, but on scale up - we had a sudden elution peak during the product load. We decided that instead of a buffer exchange, we could just dilute the protein A elution pool and it would bind to the Q. Initial capture was on protein A with citrate elution buffer. Quick story from a previous job: Purification process for a monoclonal antibody with minimal development before scale up. Unfortunately, most of the displacement chromatography I have done, was by accident with very unexpected results. I am ultimately aiming at using the resin in a displacement modeĭo you really mean "Displacement Chromatography"? Such as eluting with a charged polymer like dextran sulfate or a highly charged ion (or do you mean "flow through mode")? Displacement Chromatography is very interesting, and I have played with it a little. ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |