Single Pass Diafiltration Integrated into a Fully Continuous mAb Purification Process

Abstract

The concept of continuous manufacturing has gained significant interest from the biopharmaceutical industry over the past several years. Benefits include increased manufacturing productivity, improved quality control, reduction in plant footprint, and more flexible management of facility capacity. There are several technologies currently available that enable continuous processing for chromatography and ultrafiltration. However, a single pass diafiltration design that meets the required small molecule clearance and has been integrated into a fully continuous monoclonal antibody purification process has not been previously published. Here, the theory and design of a 3-stage single pass diafiltration step is presented. Buffer exchange greater than 99.75% was experimentally demonstrated. Several critical design aspects were incorporated to minimize system complexity and reduce the buffer volume requirements. Lastly, single pass diafiltration was demonstrated in a pilot scale continuous process with uninterrupted flow from the bioreactor through the formulation step. This work illustrates the feasibility of incorporating a single pass diafiltration step into an end-to-end continuous protein purification process. This article is protected by copyright. All rights reserved.

Continuous integrated antibody precipitation with two‐stage tangential flow microfiltration enables constant mass flow

“…The process described here had several advantages over other methods. First, it incorporates a simple, robust setup of readily available instrumentation, because TFF is a standard unit operation currently used in the pharmaceutical industry for buffer exchanges, perfusion systems, and other operations (Rucker‐Pezzini et al, ). Second, precipitation is, by nature, a concentration‐independent process; the addition of precipitant only depends on the volume that must be processed, not on the specific antibody concentration in the feed stream.…”

“…However, that strategy interrupts the fully continuous operation, and the antibody must be harvested in cycles. Another possibility is single‐pass tangential flow filtration (TFF), which allows the solid to be harvested in a single pass through a cascade of membrane filters (Casey, Gallos, Alekseev, Ayturk, & Pearl, ; Rucker‐Pezzini et al, ). The third choice is counter‐current TFF.…”

Progression of continuous downstream processing of monoclonal antibodies: Current trends and challenges

“…Single‐pass TFF (SP‐TFF) comes as an attractive alternative to traditional TFF system by using a unique flow path design and eliminating the recirculation loop (Ayturk & Marshall, ). Recently, Rucker‐Pezzini et al () addressed a major gap in the literature by demonstrating continuous diafiltration by integrating SP‐TFF into a fully continuous pilot scale production train to process 1 kg mAb in 4 days. The study reported three identical stages of diafiltration in series to achieve 99.75% buffer exchange at a feed concentration of 15 g/L and final product concentration of 148.5 g/L.…”

“…The study reported three identical stages of diafiltration in series to achieve 99.75% buffer exchange at a feed concentration of 15 g/L and final product concentration of 148.5 g/L. The current limitations on the availability of different size SP‐TFF membrane is a bottleneck in using this technology for clinical and commercial scale process (Rucker‐Pezzini et al, ). The impact of product concentration, formulation buffer type, pH, conductivity, excipient level, and process temperature on formulation offset required to counter Donnan effect in SP‐TFF is another important knowledge gap in the literature.…”

Continuous ultrafiltration/diafiltration using a 3D‐printed two membrane single pass module

“…SPTFF also proved useful for decoupling upstream and downstream process units by the inline concentration of clarified cell culture broth (Arunkumar, Singh, Peck, Borys, & Li, ; Brinkmann, Elouafiq, Pieracci, & Westoby, ). Recently, SPTFF has been reported as an interesting tool for continuous DF (Jungbauer, ; Rucker‐Pezzini et al, ). In this operation mode, several SPTFF units are sequentially connected while the DF buffer is added between the units.…”