(336d) Effects of Histidine and Sucrose on the Physical Properties and Ultrafiltration Performance of a Monoclonal Antibody

Effects of histidine
and sucrose on the physical properties and ultrafiltration performance of a
monoclonal antibody

2016 AIChE
Annual Meeting

Food, Pharmaceutical &
Bioengineering Division

Youngbin
Baek¹, Nripen Singh², Abhiram Arunkumar², and Andrew Zydney¹

¹The Pennsylvania
State University, University Park, PA 16802

²Bristol-Myers
Squibb, Global Manufacturing and Supply, Devens, MA
01434

Abstract

Monoclonal
antibodies (mAb) are currently the dominant class of
bio-therapeutics, with important applications in the treatment of cancer and
autoimmune disorders. Monoclonal antibodies are concentrated / formulated by ultrafiltration
(UF) and diafiltration (DF) to produce the formulated drug substance. There can
be significant challenges in the production of the very highly concentrated solutions
(often >200 g/L) needed to deliver the appropriate therapeutic dosage by
subcutaneous injection [1]. The objective of this study was to examine the
effects of buffer conditions on the physical characteristics and UF/DF
performance of a mAb provided by Bristol-Myers Squibb.
The mAb was characterized by its hydrodynamic radius
(determined by dynamic light scattering), zeta potential, osmotic pressure, and
viscosity, using buffers with different pH, histidine concentration, and
sucrose (used as an excipient). UF performance was evaluated using both Pellicon 3 (C- and D-screen) and Pall Omega T-series tangential
flow filtration cassettes. The hydrodynamic radius was a strong function of
both the histidine and sucrose concentrations, likely reflecting the complex
interactions between these species and the mAb /
water [2,3]. For example, the hydrodynamic radius decreased from 6.4 to 5.6 nm as
the histidine concentration increased from 10 to 250 mM,
while the addition of sucrose caused a slight increase in the mAb radius. These changes in radius were correlated with
changes in both the osmotic pressure and viscosity of the mAb
solution, particularly in the presence of high concentrations of sucrose. The change
in buffer conditions also affected the UF behavior, with the increase in
viscosity causing a significant decrease in filtrate flux (at high mAb concentrations) and maximum achievable mAb concentration. These results provide important insights
into the factors controlling the UF behavior of mAb
products, including the impact of buffer conditions on the flux and maximum
achievable protein concentration.

References

[1] Binabaji E, Ma J, Zydney AL. Intermolecular interactions
and the viscosity of highly concentrated monoclonal antibody solutions. Pharm Res. 2015:32:3102-3109.

[2] Liad SM, Du QS, Meng JZ, Pang ZW,
Huang RB. The multiple roles of histidine in protein interactions. Chem Cent J. 2013:7:44-44

[3] Nain AK, Pal
R, Sharma RK. Physicochemical study of solute–solute and solute–solvent
interactions of l-histidine in water+sucrose
solutions at different temperatures. J Mol Liq. 2012:165:154-160