High throughput screening of particle conditioning operations: I. System design and method development

Abstract

The biotech industry is under increasing pressure to decrease both time to market and development costs. Simultaneously, regulators are expecting increased process understanding. High throughput process development (HTPD) employs small volumes, parallel processing, and high throughput analytics to reduce development costs and speed the development of novel therapeutics. As such, HTPD is increasingly viewed as integral to improving developmental productivity and deepening process understanding. Particle conditioning steps such as precipitation and flocculation may be used to aid the recovery and purification of biological products. In this first part of two articles, we describe an ultra scale-down system (USD) for high throughput particle conditioning (HTPC) composed of off-the-shelf components. The apparatus is comprised of a temperature-controlled microplate with magnetically driven stirrers and integrated with a Tecan liquid handling robot. With this system, 96 individual reaction conditions can be evaluated in parallel, including downstream centrifugal clarification. A comprehensive suite of high throughput analytics enables measurement of product titer, product quality, impurity clearance, clarification efficiency, and particle characterization. HTPC at the 1 mL scale was evaluated with fermentation broth containing a vaccine polysaccharide. The response profile was compared with the Pilot-scale performance of a non-geometrically similar, 3 L reactor. An engineering characterization of the reactors and scale-up context examines theoretical considerations for comparing this USD system with larger scale stirred reactors. In the second paper, we will explore application of this system to industrially relevant vaccines and test different scale-up heuristics.

High throughput screening of particle conditioning operations: II. Evaluation of scale‐up heuristics with prokaryotically expressed polysaccharide vaccines

“…The HTPC system and procedures described in the first paper in this series were employed except where noted (Noyes et al, ). Broth was added to the Pilot‐scale vessel or microplate (2 mL, deep, square‐well, polypropylene plates; 7701–5200; Whatman, Maidstone, UK) and a steady state mixing pattern established through sustained agitation.…”

“…The configuration specified in Table : C was employed and followed the description provided in the previous publication (Noyes et al, ). Full factorial HTPC screens with pH and floc agent concentration as factors were run with lysate from two different staphylococcal CPS, SA1 and SA2, with a final volume of 1.2 mL during floc aging.…”

A modular approach for the ultra-scale-down of depth filtration

“…pH, conductivity, etc.). Screening of filtration conditions could be integrated with existing high throughput harvest, particle conditioning, and clarification workflows [3,[10][11][12][13][14][15][16]. Laboratory qualification of clinical production processes could be expedited and concluded with less material.…”

Development of a high‐throughput microscale cell disruption platform for Pichia pastoris in rapid bioprocess design

“…Scale‐down bioprocesses form the backbone of rapid bioprocess design, allowing for cost‐effective and rapid development of biopharmaceutical production processes. Specifically, small‐scale fermentation process design combined with multivariate data analysis techniques and subsequent process upscaling has proven to be effective for characterizing and optimizing industrial‐scale operations …”

“…Specifically, small-scale fermentation process design combined with multivariate data analysis techniques and subsequent process upscaling has proven to be effective for characterizing and optimizing industrial-scale operations. [1][2][3][4] Pichia pastoris, recently reassigned to the genus Komagataella, is a methanol assimilating yeast that has been receiving increased industrial attention over the past two decades due to its potential to produce a wide array of complex biopharmaceuticals. [5][6][7][8][9][10][11][12] P. pastoris offers the potential of heterologous protein secretion, however depending on physical and chemical properties of the protein of interest, this is not always a viable option.…”