Automated Purification of Antibodies Using ÄKTA Chromatography Systems

Chapter 5, Extracted from Affinity Chromatography Vol. 1: Antibodies, GE Healthcare, 2016
 

Antibodies are needed for research and industrial purposes in different quantities, from microgram to kilogram scale. It is important to design and use a purification method that will yield protein of a quality and quantity that is adequate for the particular application. The number of samples to be purified is also an important consideration. For many applications, investment in a chromatography system can save valuable time, effort and sample. Manual purification techniques are discussed in Chapter 3.

A chromatography system should be used when reproducible results are important and when manual purification becomes too time-consuming and inefficient. This can be the case when processes have to be repeated in order to obtain enough purified sample, when large sample volumes have to be handled, or when there are many different samples to be purified. Chromatography systems give more reproducible results compared with manual purification, and the progress of the purification can be monitored automatically. In addition to simple step-gradient elution, high-resolution separations with accurately controlled linear-gradient elution can be performed. Systems are robust and convenient to use and can fully utilize the high flow rates that modern chromatography media can withstand. The use of ÄKTA chromatography systems for purification of antibodies is described below.

ÄKTA start (Figure 5.1) is a cost-effective, easy-to-learn system. Together with the appropriate columns, antibodies can be purified in milligram scale in a single chromatography step with push-button control. The system includes template methods for purification of antibodies using different prepacked columns. Recovery is often better than when the same purification is performed manually. With prepacked columns and optimized purification protocols, yields and purity are highly consistent.

AKTA start

Figure 5.1. ÄKTA start.

 

AKTA avantAKTAxpress single (left) and AKTA pure chromatography systems.

Figure 5.3. ÄKTAxpress single (left) and ÄKTA pure chromatography systems.

 

Table 5.1. Standard ÄKTA protein purification systems

standard AKTA protein purification systems

Materials

     
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