Advanced multivariate data analysis to determine the root cause of trisulfide bond formation in a novel antibody–peptide fusion

Abstract

Product quality heterogeneities, such as a trisulfide bond (TSB) formation, can be influenced by multiple interacting process parameters. Identifying their root cause is a major challenge in biopharmaceutical production. To address this issue, this paper describes the novel application of advanced multivariate data analysis (MVDA) techniques to identify the process parameters influencing TSB formation in a novel recombinant antibody–peptide fusion expressed in mammalian cell culture. The screening dataset was generated with a high‐throughput (HT) micro‐bioreactor system (AmbrTM 15) using a design of experiments (DoE) approach. The complex dataset was firstly analyzed through the development of a multiple linear regression model focusing solely on the DoE inputs and identified the temperature, pH and initial nutrient feed day as important process parameters influencing this quality attribute. To further scrutinize the dataset, a partial least squares model was subsequently built incorporating both on‐line and off‐line process parameters and enabled accurate predictions of the TSB concentration at harvest. Process parameters identified by the models to promote and suppress TSB formation were implemented on five 7 L bioreactors and the resultant TSB concentrations were comparable to the model predictions. This study demonstrates the ability of MVDA to enable predictions of the key performance drivers influencing TSB formation that are valid also upon scale‐up. Biotechnol. Bioeng. 2017;114: 2222–2234. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

Red cell manufacturing using parallel stirred‐tank bioreactors at the final stages of differentiation enhances reticulocyte maturation

“…The rpms of 300, 400, 500, and 600 are equivalent to impeller tip speeds of 0.18, 0.24, 0.30, and 0.36 m/s, respectively, in the ambr® bioreactor. More information is available at the manufacturer's website and articles (Bresnick et al, 2018; Goldrick et al, 2017; Sewell et al, 2019). Viable cell density (VCD), viability, and cell diameter were measured using a Vi‐Cell XR (Beckman Coulter), which measures 4000–5500 cells per each counting using a trypan blue dye exclusion method.…”

On‐Line Control of Glucose Concentration in High‐Yielding Mammalian Cell Cultures Enabled Through Oxygen Transfer Rate Measurements

“…Micro‐scale experiments were performed on the Ambr TM ‐15 system (TAP Biosystems, Greenville, DE) that was operated as described in Ref. with 24 single vessels split into two separate culture stations where each vessel was operated with a 11–15 mL working volume. Laboratory‐scale experiments were performed in 7 L bioreactors (Applikon Biotechnology, Schiedam, The Netherlands) with a working volume of approximately 5 L. Large‐scale experiments were performed at the 50 L bioreactor scale (Applikon Biotechnology).…”

Advanced control strategies for bioprocess chromatography: Challenges and opportunities for intensified processes and next generation products

“…Methods for constructing and optimising PCA and PLS models can be found in the comprehensive review by Rolinger at al. [21], and elsewhere in the literature [52][53][54][55].…”