Chromatography affinity, immobilized monoclonal

Donohue-Rolfe, Arthur, David W.K. Acheson, Anne V. Kane, and Gerald T. Keusch. "Purification of Shiga Toxin and Shiga-Like Toxins I and II by Receptor Analog Affinity Chromatography with Immobilized PI Glycoprotein and Production of Cross-Reactive Monoclonal Antibodies." Infection and Immunity 57 (December 1989) 3888-893. 

Fig. 1 Analysis of the oligosaccharide (Glc)4 in (a) human urine and (b) serum by weak affinity chromatography. The affinity column was 5 mm LD. x 100 mm and contained immobilized monoclonal antibodies against (Glc)4. Elution was carried out under isocratic conditions at a flow rate of 0.2 mL/min using a pH 7.5, 0.02 M phosphate buffer that contained 0.1 M sodium sulfate. (From Ref. [1].)...

Proteins are frequently powerful immunogens and the availability of specific antibodies, particularly monoclonal antibodies, makes the technique of affinity chromatography very useful in the separation and purification of individual proteins. The technique has been used to purify a wide range of proteins such as hormones, membrane receptors and complement proteins. However, it is not restricted to proteins and is potentially applicable to any immunogenic substance. The availability of suitable antibodies is essential and these may be raised by whole animal polyclonal techniques or by monoclonal cell culture. The former antibodies may need some prior purification before being immobilized. 

Serpa G, Augusto EFP, Tamashiro WMSC, Ribeiro MB, Miranda EA, Bueno SMA (2005), Evaluation of immobilized metal membrane affinity chromatography for purification of IgGl monoclonal antibody, J. Chromatogr. B 816 259-268. 

Immunoaffinity separation chromatography can also be performed using immobilized specific antibodies to separate immunoglobulins. Actually immunoglobulins are antigenic molecules and, as such, they can produce specific antibodies in animals. The use of these specific antibodies as immobilized affinity ligands is the second aspect of immunoaffinity chromatography and constitutes an effective specific way to purify monoclonal and polyclonal antibodies. 

Hale, J. E., and Beidler, D. E. (1994). Purification of humanized murine and murine monoclonal antibodies using immobilized metal-affinity chromatography. Anal. Biochem. 222, 29-33. 

Where sufficient antigen is available, e.g., a synthetic peptide, the monoclonal antibodies can be purified easily and quickly by affinity chromatography. Alternatively, purified preparations of monoclonal (e.g., MARK-1, ref. 5) or polyclonal antibodies to rat or mouse F(ab )2 can be used following their immobilization to crosslinked agarose or polyacrylamide bead supports. 

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