Quasi-Continuous Production and Separation of Lysozyme Crystals on an Integrated Laboratory Plant

Timo Dobler,Marco Gleiss,Benjamin Radel,Hermann Nirschl

doi:10.3390/cryst11060713

Abstract

Vacuum crystallization with subsequent solid–liquid separation is a suitable method to produce and separate the temperature-sensitive protein lysozyme. The conventional process is performed batch-wise and on different devices, which in turn leads to disadvantages in terms of energy efficiency, contamination risk and process control. This publication therefore focuses on the application of the previously multistage process to a quasi-continuous, integrated single plant. The transfer occurs successively and starts with the substitution of the batch vessel by a process chamber. Afterwards, the filtration scale is increased and the formerly deployed membrane is replaced by an industrial filter cloth. Based on the results of these experiments, the complete process chain is successfully transferred to an integrated laboratory plant.

Highlights

Bulk protein crystallization for preparation or formulation requires new and adapted approaches in the field of process engineering and in the pharmaceutical-chemical industry
For the production or purification of temperature-sensitive materials, a vacuum crystallization followed by a solid–liquid separation is commonly suitable
In a first step of the successive transfer from a multi-stage to an integrated process, the batch vessel used for conventional preparation is replaced by a process chamber that enables the production of particles with almost equal size characteristics as the original one

Summary

Introduction

Bulk protein crystallization for preparation or formulation requires new and adapted approaches in the field of process engineering and in the pharmaceutical-chemical industry. Hekmat [1] discussed the use of large scale protein crystallization as a replacement for chromatography and formulation steps, which reduce the number of downstream processing unit operations and costs. Protein crystals, have lower mechanical strength compared to conventional ones, which makes further handling of the suspension more difficult [2]. To tackle these challenges, researchers alter proteins to enhance the crystallizabilty [3] and modify the crystals to improve stability [4]

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Results

Conclusion