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Millifluidics for Chemical Synthesis and Time-resolved Mechanistic Studies

Millifluidics for Chemical Synthesis and Time-resolved Mechanistic Studies

Published on: November 27, 2013

A versatile ceramic capillary membrane reactor system for continuous enzyme-catalyzed hydrolysis.

Lorn Messner¹, Marieke H Antink², Tongwei Guo²

¹Institute for Technical Chemistry Leibniz University Hannover Hannover Germany.

Engineering in Life Sciences

|September 29, 2021

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A new continuous ceramic capillary membrane reactor system (CCCMRS) offers stable, reproducible, and controllable enzyme-catalyzed hydrolysis. This system demonstrates long-term stability for antibody fragment production and peptide generation from protein isolates.

Area of Science:

Biotechnology
Chemical Engineering
Enzyme Technology

Background:

Classical batch processes for enzyme-catalyzed hydrolysis have limitations in stability and reproducibility.
Continuous processing offers potential advantages for enzymatic reactions.

Purpose of the Study:

To develop and evaluate a continuous ceramic capillary membrane reactor system (CCCMRS) for enzyme-catalyzed hydrolysis.
To demonstrate the long-term stability and applicability of the CCCMRS for producing antibody fragments and peptides.

Main Methods:

Development of a CCCMRS with immobilized proteolytic enzymes on ceramic capillary membranes.
Long-term stability studies of the CCCMRS using two applications: human IgG cleavage and protein isolate digestion.
Analysis of reaction products using Size Exclusion-High Performance Liquid Chromatography (SEC-HPLC) and Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC).

Keywords:

antibody fragments bioactive peptides food proteins immobilized enzymes proteolysis

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Main Results:

The CCCMRS demonstrated long-term stability over 33.3 hours for human IgG cleavage into Fab and Fc fragments.
Continuous digestion of pea and almond protein isolates yielded a constant peptide fingerprint over 45 hours.
The system showed stability, reproducibility, and controllability, adaptable to various enzymes and conditions.

Conclusions:

The developed CCCMRS is a robust platform for continuous enzyme-catalyzed hydrolysis.
The system enables efficient and stable production of valuable biomolecules like antibody fragments and peptides.
The CCCMRS facilitates further bioactivity studies by providing consistent peptide generation.