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Shear contributions to cell culture performance and product recovery in ATF and TFF perfusion systems

Course Orientation and Learning Objectives

1. Introduction

Foundations of Shear in ATF and TFF Perfusion

2. Materials and methods

Quantifying Shear and Cell Damage: Methods and Metrics

Cell Culture Setup for Shear Studies

Measuring Cell Death and Lysis under Shear

Offline Modeling of Product Sieving under Shear

Analytical Methods for Product Quality under Shear

Statistics for Shear-Impact Experiments

3. Results

Comparative Shear Exposure: ATF versus TFF

Debris and Particle Size Effects on Product Sieving

Root Causes of Lysis in TFF Loops

Pump-Induced Shear and Mitigation

Low-Shear Pumping Strategies to Improve Outcomes

4. Discussion

Design and Scale-Up Framework to Control Shear in ATF and TFF

5. Conclusions

Key Takeaways and Implementation Checklist

Knowledge Checks

Quiz: Hydrodynamic Parameters and Shear Metrics

Quiz: ATF vs. TFF Shear Impacts on Cells and Product

Quiz: Pumps, Lysis Mechanisms, and Mitigations

Quiz: Product Sieving, Fouling, and Particle Distributions

Quiz: Scale-Up and Control Strategy for Shear Management

Course Feedback

Survey: Learner Confidence and Applicability to Processes

Shear Management in ATF and TFF Perfusion: Diagnosis to Scale-Up is a practical, evidence-driven course for bioprocess scientists and engineers who run high-density perfusion with alternating tangential flow (ATF) or tangential flow filtration (TFF) cell-retention. You will learn how hydrodynamic forces—shear stress, energy dissipation rate, and extensional/turbulent shear—shape cell health, viability, specific productivity, and product quality. Through a structured journey from fundamentals to scale-up, you will master how crossflow, recirculation, transmembrane pressure (TMP), pulsation, and pump selection govern shear exposure in real systems. The program translates analytics (viability, apoptosis/necrosis, LDH, PSD), membrane performance (sieving, fouling resistance, permeability), and statistical design into clear operating envelopes and control strategies. Learn why peristaltic pumps often drive lysis in TFF loops, how low-shear centrifugal options can dramatically recover product sieving and viability, and how to set robust, scalable ranges for crossflow and TMP. By the end, you will have a ready-to-use checklist to diagnose shear problems, tune parameters without sacrificing productivity, and de-risk process transfer from lab to pilot and commercial scales.

Shear Management in ATF and TFF Perfusion: Diagnosis to Scale-Up

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            <p>Shear Management in ATF and TFF Perfusion: Diagnosis to Scale-Up equips process development teams to quantify, troubleshoot, and control hydrodynamic stresses that govern cell performance and product recovery across perfusion architectures. The course begins by establishing a common vocabulary for shear stress and energy dissipation rate, and by distinguishing extensional, turbulent, and pulsation-driven shear in ATF and TFF loops. Building from these foundations, learners connect hydrodynamic drivers—crossflow velocity, recirculation rate, transmembrane pressure, and oscillation profiles—to phenotypic outcomes including viability, lysis, specific productivity, and product quality attributes such as aggregation and fragmentation. Throughout, a structured reference framework links course goals to practical tasks: diagnosing shear sources, prioritizing mitigations, and translating findings into robust operating envelopes.</p>
<p>Materials and methods modules provide a rigorous toolkit to measure shear proxies (pressure, flow, TMP ripple, ATF amplitude/frequency, TFF crossflow), assess cell damage (viability, apoptosis/necrosis, LDH, PSD), and characterize sieving, fouling resistance, and permeability with attention to data quality, replication, and statistical power. Results synthesize comparative evidence of shear exposure in ATF versus TFF, identify root causes of elevated lysis in TFF—particularly peristaltic pump–induced extensional and squeeze shear—and demonstrate how low-shear centrifugal pumping can recover viability and product passage while maintaining productivity through re-optimization of crossflow and TMP. The discussion culminates in a scale-up framework that integrates first-principles and data-driven modeling to set control strategies for pump selection, pulsation management, and hold-up minimization from lab to commercial scale. Knowledge checks reinforce key concepts, and a closing checklist supports implementation of a lifecycle risk-based approach to shear management in perfusion processes.</p>
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