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Membrane-Based Hybrid Method for Purifying PEGylated Proteins

PEGylated proteins are usually purified using chromatographic methods, which are limited in terms of both speed and scalability. In this paper, we describe a microfiltration membrane-based hybrid method for purifying PEGylated proteins. Polyethylene glycol (or PEG) is a lower critical solution tempe...

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Detalles Bibliográficos
Autores principales: Lam, Shing Fung, Shang, Xiaojiao, Ghosh, Raja
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9966431/
https://www.ncbi.nlm.nih.gov/pubmed/36837684
http://dx.doi.org/10.3390/membranes13020182
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author Lam, Shing Fung
Shang, Xiaojiao
Ghosh, Raja
author_facet Lam, Shing Fung
Shang, Xiaojiao
Ghosh, Raja
author_sort Lam, Shing Fung
collection PubMed
description PEGylated proteins are usually purified using chromatographic methods, which are limited in terms of both speed and scalability. In this paper, we describe a microfiltration membrane-based hybrid method for purifying PEGylated proteins. Polyethylene glycol (or PEG) is a lower critical solution temperature polymer which undergoes phase transition in the presence of a lyotropic salt and forms micelle-like structures which are several microns in size. In the proposed hybrid method, the PEGylated proteins are first converted to their micellar form by the addition of a lyotropic salt (1.65 M ammonium sulfate). While the micelles are retained using a microfiltration membrane, soluble impurities such as the unmodified protein are washed out through the membrane. The PEGylated proteins thus retained by the membrane are recovered by solubilizing them by removing the lyotropic salt. Further, by precisely controlling the salt removal, the different PEGylated forms of the protein, i.e., mono-PEGylated and di-PEGylated forms, are fractionated from each other. Hybrid separation using two different types of microfiltration membrane devices, i.e., a stirred cell and a tangential flow filtration device, are examined in this paper. The membrane-based hybrid method for purifying PEGylated proteins is both fast and scalable.
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spelling pubmed-99664312023-02-26 Membrane-Based Hybrid Method for Purifying PEGylated Proteins Lam, Shing Fung Shang, Xiaojiao Ghosh, Raja Membranes (Basel) Article PEGylated proteins are usually purified using chromatographic methods, which are limited in terms of both speed and scalability. In this paper, we describe a microfiltration membrane-based hybrid method for purifying PEGylated proteins. Polyethylene glycol (or PEG) is a lower critical solution temperature polymer which undergoes phase transition in the presence of a lyotropic salt and forms micelle-like structures which are several microns in size. In the proposed hybrid method, the PEGylated proteins are first converted to their micellar form by the addition of a lyotropic salt (1.65 M ammonium sulfate). While the micelles are retained using a microfiltration membrane, soluble impurities such as the unmodified protein are washed out through the membrane. The PEGylated proteins thus retained by the membrane are recovered by solubilizing them by removing the lyotropic salt. Further, by precisely controlling the salt removal, the different PEGylated forms of the protein, i.e., mono-PEGylated and di-PEGylated forms, are fractionated from each other. Hybrid separation using two different types of microfiltration membrane devices, i.e., a stirred cell and a tangential flow filtration device, are examined in this paper. The membrane-based hybrid method for purifying PEGylated proteins is both fast and scalable. MDPI 2023-02-02 /pmc/articles/PMC9966431/ /pubmed/36837684 http://dx.doi.org/10.3390/membranes13020182 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lam, Shing Fung
Shang, Xiaojiao
Ghosh, Raja
Membrane-Based Hybrid Method for Purifying PEGylated Proteins
title Membrane-Based Hybrid Method for Purifying PEGylated Proteins
title_full Membrane-Based Hybrid Method for Purifying PEGylated Proteins
title_fullStr Membrane-Based Hybrid Method for Purifying PEGylated Proteins
title_full_unstemmed Membrane-Based Hybrid Method for Purifying PEGylated Proteins
title_short Membrane-Based Hybrid Method for Purifying PEGylated Proteins
title_sort membrane-based hybrid method for purifying pegylated proteins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9966431/
https://www.ncbi.nlm.nih.gov/pubmed/36837684
http://dx.doi.org/10.3390/membranes13020182
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