High concentration tangential flow ultrafiltration of stable monoclonal antibody solutions with low viscosities

Abstract

During production of concentrated monoclonal antibody formulations by tangential flow ultrafiltration (TFF), high viscosities and aggregation often cause extensive membrane fouling, flux decay and low product yields. To address these challenges, the co-solutes histidine or imidazole were added at high concentrations from 250 to 320 mM to reduce the viscosity by up to tenfold relative to conventional low co-solute formulations, to as low as 40 cP at 250 mg/mL. At high mAb concentrations of up to 280 mg/mL, the transmembrane flux was increased threefold by adding high concentrations of co-solutes that also lowered the viscosity. Furthermore, the co-solutes also increased the mAb gel point concentration c g by up to 100 mg/mL mAb and thus enhanced concentration polarization-driven back-diffusion of the mAb at the membrane wall, which led to increased fluxes. The low viscosity and hollow fiber filter modules with straight flow paths enabled more uniform TMP and wall shear stress τ w profiles, which mitigated the reversible flux decay that results from an axial decline in the local TMP. The concentrated mAb was stable by SEC before and after extended storage at 4°C and 37°C.

Ultrafiltration behavior of monoclonal antibodies and Fc‐fusion proteins: Effects of physical properties

“…This behavior is a direct result of the back‐filtration that occurs near the cassette exit due to the high pressure losses associated with flow of the concentrated protein solution through the screened channel. Reductions in the protein solution viscosity lead to an increase in the maximum achievable protein concentration ( C max ), consistent with results from previous experimental studies (Goswami et al, ; Hung et al, ).…”

“…The development of the ultrafiltration (UF) step in the platform process can be more challenging since there are currently no high throughput approaches that effectively capture the behavior of tangential flow filtration (TFF) systems and there is a lack of quantitative understanding of the key physical properties that control the filtrate flux and UF behavior for these antibody‐based products. Goswami et al () found that the large viscosity of highly concentrated protein solutions increases the back pressure of the pump and the processing time during UF, while Hung et al () showed that the filtrate flux in UF could be increased using excipients like histidine or imidazole that reduce the viscosity, although no quantitative relationships between the viscosity and the filtrate flux were presented. More recently, Binabaji et al () developed a model for the filtrate flux during UF of highly concentrated mAb solutions that incorporates the effects of the solution viscosity and protein osmotic pressure on the flux; this model is discussed in more detail subsequently.…”

Enhancing the Injectability of High Concentration Drug Formulations Using Core Annular Flows

“…This figure reports the injection force (for a flow rate of 4 mL min −1 through a 27G needle) as a function of concentration for four monoclonal antibody solutions of the IgG1 isotype that have been reported in literature. [10][11][12][13] This figure also highlights the fact that an extensive range of formulation concentrations require more than 50 N to be injected-the average maximum force that can be applied in a pinching motion to a syringe for subcutaneous injections. [14][15][16] While larger needle gauges or prolonged injection times through the use of syringe pumps could overcome challenges associated with the low injectability of high concentration formulations, these approaches would negate the benefits offered by subcutaneous delivery due to a larger degree of pain and the requirement of a hospital setting, respectively.…”

“…[8,9] A nonlinear relationship between formulation concentration and viscosity makes subcutaneous formulations very viscous (20-60 cP) and therefore harder to inject, as illustrated in Figure 1a: a highly viscous fluid (dyed blue) spreads significantly less than a low viscosity fluid (dyed green), when injected in a sponge at the maximal force we could apply manually (around 50 N). [10][11][12][13] Consequently, the applicable force sets a limit to the concentrations of current formulations, as shown in Figure 1b. This figure reports the injection force (for a flow rate of 4 mL min −1 through a 27G needle) as a function of concentration for four monoclonal antibody solutions of the IgG1 isotype that have been reported in literature.…”

“…b) The force required to manually inject four (labeled 1-4) highly concentrated monoclonal antibody solutions of the IgG1 isotype reported in literature exceeds the maximum force that is applicable in a pinching motion (50 N). [10][11][12][13][14][15][16] (Forces calculated for a flow rate of 4 mL min −1 ). c) Schematic cross-sections of unlubricated flow and core annular flow through a needle.…”