Monoclonal Antibody purification

This method of separation has been applied to monoclonal antibody purification (A. Schwarz, personal communication). The selectivity for the antibodies is played by the choice of the ligand while the HCIC effect is still the one described above. 

In monoclonal antibody purification, biological risks are primarily related to the host animal cells, but also to animal supplements for culture medium such as fetal bovine serum or pure proteins (e.g., bovine albumin, insulin, and transferrin). A special risk associated with production of antibodies with rodent cell lines is their high load of C-type particles. These particles are considered as incomplete retroviruses. The danger regarding infecting humans is not clear. Thus, the efficient separation of these particles must be guaranteed. These particles are quantified either by immunological techniques or electron microscopy. 

For such validation studies, one of the critical factors is the choice of viruses. This will unavoidably be dependent on the starting raw material in the case of monoclonal antibody purification, which is frequently done from rodent cell line cultures, retroviruses are of particular concern. 

Lihme, A., and Bendix Hansen, M. (1997). Protein A mimetic for large scale monoclonal antibody purification. Biotechnol. Lab. 15, 30-31. 

Hubbard, B. 2005. Platform approaches to monoclonal antibody purification. IBC Conference Antibody Production Downstream Processing, Mar 8-11, San Diego, CA. 

STREAMLINE expanded bed adsorption is particularly suited for large scale recombinant protein and monoclonal antibody purification. STREAMLINE adsorbents are specially designed for use in STREAMLINE columns. The technique requires no sample clean up and so combines sample preparation and capture in a single step. As shown in Figure 10, crude sample is applied to an expanded bed of STREAMLINE media, target proteins are captured whilst cell debris, particulate matter, whole cells, and contaminants pass through. Flow is reversed and the target protein is desorbed in the elution buffer. 

The objective of this work was to produce an efficient, routine procedure for monoclonal antibody purification. 

For large scale recombinant protein and monoclonal antibody purification. The technique requires no sample clean up and enables clarification, concentration and capture in a single step. 

Jungbauer, A. "Scaleup of monoclonal antibody purification using radial streaming chromatography" Biotech, and Bioeng. 1988, 32, pp 326-333. 

Factor VIII concentrates can be classified according to their level of purity, which refers to the specific activity of factor VIII in the product. Cryoprecipitate is a low-purity product, containing a specific factor VIII activity level of less than 5 units/mg of protein. This product is no longer considered a primary treatment for factor VIE deficiency in countries where factor VEI concentrates are available, because cryoprecipitate does not undergo a viral inactivation process. Intermediate-purity products have a specific activity of factor VIE of 1 to 10 units/mg of protein, while high-purity products have a specific activity of 50 to 1,000 units/mg of protein. Ultrahigh-purity plasma-derived products are prepared with monoclonal antibody purification steps and have a specific activity of 3,000 units/mg of protein prior to the addition of albumin as a stabihzer. 

H. J., A base-tolerant resin for Monoclonal Antibody purification. 3rd International Symposium on Downstream Processing of Genetically Engineered Antibodies and Related Molecules, 2004, Nice, France. 

Example 9.14 Monoclonal Antibody Purification by Affinity Adsorption... 

Aldington, S. and Bonnerjea, J. (2007) Scale-up of monoclonal antibody purification processes. J. Chromatosr- B, 848, 64-78. 

Grandic, P. (1994) Monoclonal antibody purification guide. art3. American Biotech. Lab. 12(8), 16, 18. 

Boi C. Membrane adsorbers as purification tools for monoclonal antibody purification. /. Chromatogr. B 2007 848 19-27. 

Pharmacia Biotech. (1997). Monoclonal antibody purification. Technical handbook. Cat. No. 18-1037-46. Pharmacia, Uppsala. 

Kelley, B. (2007) Very large scale monoclonal antibody purification the... 

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