Purification of Antibodies by Liquid Chromatography

Liquid chromatography is very often the method of choice for adsorption, separation, and purification of antibodies. Chromatographic separations are based on the differential adsorption and migration speed of components of a protein mixture through a column filled with small particles called chromatographic sorbent or solid phase.71 At the level of adsorption, solid phases are 

TABLE 4 Yields and Purification Factors during Purification of Monoclonal IgM 

The starting volume is 150 liters hybridoma culture supernatant Kp and K, are the purification factors related to protein or volume respectively according to Steindl et al. (  

It is beyond the scope of this chapter to give details of the technology for the separation of antibodies. Instead, first some basis is presented and then how liquid chromatography is usable for the adsorption, separation, and purification of antibodies is explained. Method development phases such as, for instance, resin screening, optimization of running conditions, flow sheeting, operational parameters for productivity optimization, and the choice of system components, will not be detailed here. For general reviews, see Refs. 71 and 72. 

All individual methodologies described in the following, such as ion exchange, hydrophobic interaction, and affinity chromatography, are potentially usable in packed-bed mode as well as in fluid-bed mode if the solid phase sorbent meets the specific requirements of density and particle size. The only exception today is gel filtration, which requires a large number of plates for an acceptable resolution that the fluid bed cannot guarantee. 

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