Monoclonal antibodies purification technology

The practical achievement of this goal was held up for 18 years, primarily because of the great difficulty in isolation and purification of single-species proteins from the immune repertoire. During that time, many attempts to elicit catalysis by inhomogeneous (i.e. polyclonal) mixtures of antibodies were made and failed (e.g. Raso and Stollar, 1975 Summers, 1983). The problem was resolved in 1976 by Kbhler and Milstein s development of hybridoma technology, which has made it possible today both to screen rapidly the complete immune repertoire and to produce in vitro relatively large amounts of one specific monoclonal antibody species (Kohler and Milstein, 1975 Kohler et al., 1976). 

Thommes J, Born C, Biselli M, Wandrey C, Kula MR (1995) Purification of monoclonal antibodies by fluidized bed adsorption. In Beuvery CE, Griffiths JB, Zeijlmaker WP (ed) Animal cell technology developments towards the 21st century. Kluwer, p 515... 

Thiophilic adsorption chromatography has been described for the purification of murine monoclonal antibodies from hybridoma cell culture containing fetal bovine serum.154 Due to the very low concentration of immunoglobulins in cell culture supernatants, binding capacity remains modest in spite of the presence of 0.5 to 1 M potassium sulfate. Further developments of this technology described by Nopper et al.155 indicated that thiophilic sorbents could be modified in their structure to increase the specificity and the binding capacity. 

The recent technological advances In molecular biology have permitted the production of a variety of Important molecules on a very large scale. In most cases the compounds of Interest are produced In a broth with a large number of similar species and require purification. Compounds to be employed for therapeutic purposes require a high degree of purity. This Is especially true If the molecule Is the product of a transformed cell, as are monoclonal antibodies. 

This chapter describes the production of anti-serums, the purification of antibodies, and the most important immunological screening technologies. For monoclonal antibodies, I refer you to Methods Enzymol. I (1986) 121, Hybridoma Technology and Monoclonal Antibodies. I also touch only briefly upon the newer in-vitro immunization methods in the following (with friendly consultation from Dr. Rose-John, Mainz). 

Due to the importance of this technology for protein expression, manufacturers Sigma, Serotec, Abeam and many others offer a wide range of products for the detection, isolation and purification of tagged proteins. These vendors also provide monoclonal and polyclonal antibodies specific for the most commonly... 

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