A stepwise mechanism for aqueous two-phase system formation in concentrated antibody solutions

Proceedings of the National Academy of Sciences of the United States of America
Bradley A RogersPaul S Cremer

Abstract

Aqueous two-phase system (ATPS) formation is the macroscopic completion of liquid-liquid phase separation (LLPS), a process by which aqueous solutions demix into 2 distinct phases. We report the temperature-dependent kinetics of ATPS formation for solutions containing a monoclonal antibody and polyethylene glycol. Measurements are made by capturing dark-field images of protein-rich droplet suspensions as a function of time along a linear temperature gradient. The rate constants for ATPS formation fall into 3 kinetically distinct categories that are directly visualized along the temperature gradient. In the metastable region, just below the phase separation temperature, T ph , ATPS formation is slow and has a large negative apparent activation energy. By contrast, ATPS formation proceeds more rapidly in the spinodal region, below the metastable temperature, T meta , and a small positive apparent activation energy is observed. These region-specific apparent activation energies suggest that ATPS formation involves 2 steps with opposite temperature dependencies. Droplet growth is the first step, which accelerates with decreasing temperature as the solution becomes increasingly supersaturated. The second step, however, involves dr...Continue Reading

References

Sep 1, 1992·Protein Science : a Publication of the Protein Society·R Bhat, S N Timasheff
Oct 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·J A ThomsonG B Benedek
Nov 19, 1985·Biochemistry·T Arakawa, S N Timasheff
Apr 1, 1971·The Journal of Pharmacy and Pharmacology·A T Florence, J A Rogers
Jan 9, 1996·Proceedings of the National Academy of Sciences of the United States of America·C LiuG B Benedek
Apr 19, 2002·Journal of the American Chemical Society·Hanbin MaoPaul S Cremer
Oct 17, 2002·Analytical Chemistry·Hanbin MaoPaul S Cremer
Oct 23, 2002·Proceedings of the National Academy of Sciences of the United States of America·Onofrio AnnunziataGeorge B Benedek
Jan 17, 2003·Proceedings of the National Academy of Sciences of the United States of America·Onofrio AnnunziataGeorge B Benedek
Dec 11, 2003·Journal of the American Chemical Society·Yanjie ZhangPaul S Cremer
Aug 25, 2004·Methods : a Companion to Methods in Enzymology·Neer Asherie
Oct 12, 2004·The Journal of Chemical Physics·Mrinal ShahPeter G Vekilov
Apr 24, 2007·Biophysical Journal·Tangir AhamedJörg Thömmes
Oct 13, 2007·Physical Review Letters·Frédéric CardinauxPeter Schurtenberger
Apr 15, 2008·Journal of Colloid and Interface Science·Luke ParkinsonJohn Ralston
May 24, 2008·Nature·Peter J LuDavid A Weitz
Apr 29, 2010·The Journal of Physical Chemistry. B·Weichun PanVassiliy Lubchenko
Feb 12, 2011·Biotechnology Progress·Rachael A LewusStanley I Sandler
Jun 4, 2011·Journal of Pharmaceutical Sciences·Bruce D MasonJifeng Zhang
Aug 12, 2009·Journal of Physics. Condensed Matter : an Institute of Physics Journal·Thomas Gibaud, Peter Schurtenberger
Sep 17, 2011·Proceedings of the National Academy of Sciences of the United States of America·Ying WangGeorge B Benedek
Nov 16, 2011·The Journal of Biological Chemistry·Ramil F LatypovDingjiang Liu
Jan 5, 2012·Lab on a Chip·Thomas KrebsRemko Boom
Apr 13, 2012·Journal of Physics. Condensed Matter : an Institute of Physics Journal·Peter G Vekilov
Aug 1, 2012·Proceedings of the National Academy of Sciences of the United States of America·Ying WangGeorge B Benedek
Oct 5, 2013·The Journal of Chemical Physics·Ying WangGeorge B Benedek

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Hereditary Sensory Autonomic Neuropathy

Hereditary Sensory Autonomic Neuropathies are a group of inherited neurodegenerative disorders characterized clinically by loss of sensation and autonomic dysfunction. Here is the latest research on these neuropathies.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Landau-Kleffner Syndrome

Landau Kleffner syndrome (LKS), also called infantile acquired aphasia, acquired epileptic aphasia, or aphasia with convulsive disorder, is a rare childhood neurological syndrome characterized by the sudden or gradual development of aphasia (the inability to understand or express language) and an abnormal electroencephalogram. Discover the latest research on LKS here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Regulation of Vocal-Motor Plasticity

Dopaminergic projections to the basal ganglia and nucleus accumbens shape the learning and plasticity of motivated behaviors across species including the regulation of vocal-motor plasticity and performance in songbirds. Discover the latest research on the regulation of vocal-motor plasticity here.