Recombinant Protein L: Production, Purification and Characterization of a Universal Binding Ligand

Stefan Kittler; Julian Ebner; Mihail Besleaga; Johan Larsbrink; Barbara Darnhofer; Ruth Birner-Gruenberger; Silvia Schobesberger; Christopher K Akhgar; Andreas Schwaighofer; Bernhard Lendl; Oliver Spadiut

doi:10.1016/j.jbiotec.2022.10.002

Recombinant Protein L: Production, Purification and Characterization of a Universal Binding Ligand

J Biotechnol. 2022 Nov 20:359:108-115. doi: 10.1016/j.jbiotec.2022.10.002. Epub 2022 Oct 4.

Affiliations

¹ Research Division Integrated Bioprocess Development, Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Gumpendorfer Strasse 1a, 1060 Vienna, Austria; Alfred Gruber GmbH, Nordstrasse 6, 5301 Eugendorf, Austria. Electronic address: stefan.kittler@tuwien.ac.at.
² Research Division Integrated Bioprocess Development, Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Gumpendorfer Strasse 1a, 1060 Vienna, Austria; Alfred Gruber GmbH, Nordstrasse 6, 5301 Eugendorf, Austria. Electronic address: julian.ebner@tuwien.ac.at.
³ Research Division Integrated Bioprocess Development, Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Gumpendorfer Strasse 1a, 1060 Vienna, Austria. Electronic address: mihail.besleaga@tuwien.ac.at.
⁴ Wallenberg Wood Science Center, Division of Industrial Biotechnology, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden. Electronic address: johan.larsbrink@chalmers.se.
⁵ Research Division Functional Proteomics and Metabolic Pathways, Diagnostic and Research Institute of Pathology, Medical University of Graz, Stiftingtalstrasse 24, 8010 Graz, Austria. Electronic address: b.darnhofer@medunigraz.at.
⁶ Research Division Functional Proteomics and Metabolic Pathways, Diagnostic and Research Institute of Pathology, Medical University of Graz, Stiftingtalstrasse 24, 8010 Graz, Austria; Research Division Bioanalytics, Institute of Chemical Technology and Analytics, TU Wien, Getreidemarkt 9/164, 1060 Vienna, Austria. Electronic address: ruth.birner-gruenberger@tuwien.ac.at.
⁷ Research Division Organic & Biological Chemistry, Institute of Applied Synthetic Chemistry, TU Wien, Getreidemarkt 9/163, 1060 Vienna, Austria. Electronic address: silvia.schobesberger@tuwien.ac.at.
⁸ Research Division Environmental Analytics, Process Analytics and Sensors, Institute of Chemical Technology and Analytics, TU Wien, Getreidemarkt 9/164, 1060 Vienna, Austria. Electronic address: christopher.akhgar@tuwien.ac.at.
⁹ Research Division Environmental Analytics, Process Analytics and Sensors, Institute of Chemical Technology and Analytics, TU Wien, Getreidemarkt 9/164, 1060 Vienna, Austria. Electronic address: andreas.schwaighofer@tuwien.ac.at.
¹⁰ Research Division Environmental Analytics, Process Analytics and Sensors, Institute of Chemical Technology and Analytics, TU Wien, Getreidemarkt 9/164, 1060 Vienna, Austria. Electronic address: bernhard.lendl@tuwien.ac.at.
¹¹ Research Division Integrated Bioprocess Development, Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Gumpendorfer Strasse 1a, 1060 Vienna, Austria. Electronic address: oliver.spadiut@tuwien.ac.at.

PMID: 36206851
DOI: 10.1016/j.jbiotec.2022.10.002

Abstract

Protein L (PpL) is a universal binding ligand that can be used for the detection and purification of antibodies and antibody fragments. Due to the unique interaction with immunoglobulin light chains, it differs from other affinity ligands, like protein A or G. However, due to its current higher market price, PpL is still scarce in applications. In this study, we investigated the recombinant production and purification of PpL and characterized the product in detail. We present a comprehensive roadmap for the production of the versatile protein PpL in E. coli.

Keywords: Affinity ligand; Bioreactor; E. coli; Protein L; Recombinant production.

MeSH terms

Bacterial Proteins* / metabolism
Chromatography, Affinity
Escherichia coli* / genetics
Escherichia coli* / metabolism
Immunoglobulin Fragments
Immunoglobulin Light Chains
Ligands
Protein Binding
Recombinant Proteins / metabolism

Substances

Ligands
Bacterial Proteins
Recombinant Proteins
Immunoglobulin Fragments
Immunoglobulin Light Chains