Method and apparatus for chromatographic purification

Abstract

The present invention relates to a method and an apparatus suitable for a continuous chromatography process which only needs three separation columns. The process is a two step procedure comprising two chromatographic steps. The first chromatographic step (capture) is performed alternating and sequentially on two separation columns, the second chromatographic step (polishing) is performed, also sequentially, on the third column.

Description

[0001]The present invention relates to a method and an apparatus suitable for a continuous chromatography process which only needs three separation columns. The process is a two step procedure comprising two chromatographic steps. The first chromatographic step (capture) is performed alternating and sequentially on preferably two separation columns, the second chromatographic step (polishing) is performed, also sequentially, on the third column.BACKGROUND OF THE INVENTION[0002]The large-scale, economic purification of proteins is increasingly an important problem for the biotechnology and pharmaceutical industry. Typically, proteins are produced by cell culture, using either mammalian or bacterial cell lines engineered to produce the protein of interest by insertion of a recombinant plasmid containing the gene for that protein. Since the cell lines used are living organisms, they must be fed with a complex growth medium, containing sugars, amino acids, and growth factors, usually su...

AI Technical Summary

Benefits of technology

[0036]In a preferred embodiment, the apparatus further comprises a connecting line between the fluid outlet of separation unit A1 and the fluid inlet of separation unit A2 and a connecting line between the fluid outlet of separation unit A2 and the fluid inlet of separation unit A1 (connecting lines F-F in FIGS. 2, 3 and 4), thus enabling a fluid communication between the outlet of separation unit A1 and the inlet of separation unit A2 as well as a fluid communication between the outlet of separation unit A2 and the inlet of separation unit A1. In a very preferred embodiment, at least one valve is located in the connecting line between the outlet of separation unit A1 and the inlet of separation unit A2 and at least one valve is located in the connecting line between the outlet of separation unit A2 and the inlet of separation unit A1. Typically the valves are located close to the outlets of separation units A1 and A2 and / or close to the inlets of separation units A1 and A2.

Problems solved by technology

The large-scale, economic purification of proteins is increasingly an important problem for the biotechnology and pharmaceutical industry.

Separation of the desired protein from the mixture of compounds fed to the cells and from the by-products of the cells themselves to a purity sufficient for use as a human therapeutic poses a formidable challenge.

Such disruption releases the entire contents of the cell into the homogenate, and in addition produces subcellular fragments that are difficult to remove due to their small size.

The same problem arises, although on a smaller scale, with directly secreted proteins due to the natural death of cells and release of intracellular host cell proteins in the course of the protein production run.

Despite its common use, affinity chromatography is costly, particularly at the industrial scale necessary to purify therapeutic proteins.

Simple batch chromatography technique is well accepted in the industrial applications, however this technology is expensive due to long processing times and high operation costs (e.g. large solvent amounts, expensive resins and hardware).

This technique is also sensitive to operational conditions (e.g. product titer, residence time and feeding rate (product losses starting from 80% dynamic binding capacity values).

Some alternative semi-continuous technologies were developed as well, meaning that they connect two or three different chromatography modes, but do not allow one to have a continuous feed.

Images