Phase Behavior of an Intact Monoclonal Antibody

Phase Behavior of an Intact Monoclonal Antibody

Ahamed, T.
Esteban, B.N.
Ottens, M.
van Dedem, G.W.
Van der Wielen, L.A.
Bisschops, M.A.
Lee, A.
Pham, C.
Thömmes, J.

Applied Sciences

Biotechnology

2007-04-20

Understanding protein phase behavior is important for purification, storage, and stable formulation of protein drugs in the biopharmaceutical industry. Glycoproteins, such as monoclonal antibodies (MAbs) are the most abundant biopharmaceuticals and probably the most difficult to crystallize among water-soluble proteins. This study explores the possibility of correlating osmotic second virial coefficient (B22) with the phase behavior of an intact MAb, which has so far proved impossible to crystallize. The phase diagram of the MAb is presented as a function of the concentration of different classes of precipitants, i.e., NaCl, (NH4)2SO4, and polyethylene glycol. All these precipitants show a similar behavior of decreasing solubility with increasing precipitant concentration. B22 values were also measured as a function of the concentration of the different precipitants by selfinteraction chromatography and correlated with the phase diagrams. Correlating phase diagrams with B22 data provides useful information not only for a fundamental understanding of the phase behavior of MAbs, but also for understanding the reason why certain proteins are extremely difficult to crystallize. The scaling of the phase diagram in B22 units also supports the existence of a universal phase diagram of a complex glycoprotein when it is recast in a protein interaction parameter.

http://resolver.tudelft.nl/uuid:92f4bdf6-8614-4004-8823-a95d04092787

https://doi.org/10.1529/biophysj.106.098293

Biophysical Society

1542-0086

Biophysical Journal, 93 (July) 2007

Institutional Repository

journal article

(c) 2007 Ahamed, T.; Biophysical Society