albumin - publications

Predict more albumin - ligand interactions now!


1. J Pharm Sci. 2012 Mar 12. doi: 10.1002/jps.23096. [Epub ahead of print]

Biophysical characterization and stabilization of the recombinant albumin fusion
protein sEphB4-HSA.

Shi S, Liu J, Joshi SB, Krasnoperov V, Gill P, Middaugh CR, Volkin DB.

Department of Pharmaceutical Chemistry, Macromolecule and Vaccine Stabilization
Center, University of Kansas, Lawrence, Kansas 66047.

The use of albumin fusion proteins as therapeutic drug candidates is an
attractive approach to design novel biopharmaceuticals with increased circulation
time in vivo. The purpose of this work was to characterize and stabilize the
fusion protein sEphB4-human serum albumin (HSA), a 120 kDa protein containing the
extracellular domain of EphB4 and HSA, and to identify stabilizing excipients for
storage. Comparative biophysical studies combined with empirical phase diagram
analysis show that both structural integrity and conformational stability of
sEphB4 and sEphB4-HSA are best maintained above pH 5 and below 50°C, with
different structural phases observed outside this range. A major physical
degradation pathway for sEphB4-HSA is formation of soluble aggregates. Excipient
studies using size-exclusion chromatography (SEC), differential scanning
calorimetry (DSC), and fluorescence spectroscopy identified disaccharide sugars
(e.g., sucrose and trehalose) as effective stabilizers against protein
aggregation, and NaCl as an effective stabilizer for protecting overall
conformational stability. A combination of biophysical studies with sEphB4-HSA
and its individual component proteins (sEphB4 and HSA), along with correlation
analysis of SEC and DSC stability data in the presence of different excipients
suggest that the aggregation pathway of the albumin fusion protein is primarily
mediated by the sEphB4 rather than the HSA component. © 2012 Wiley Periodicals,
Inc. and the American Pharmacists Association J Pharm Sci.

Copyright © 2012 Wiley Periodicals, Inc.

PMID: 22411527 [PubMed - as supplied by publisher]