Purification of protein therapeutics via high-affinity supramolecular host-guest interactions.

Nature Biomedical Engineering
Jaeyeon AnKimoon Kim

Abstract

Efficient purification is crucial to providing large quantities of recombinant therapeutic proteins, such as monoclonal antibodies and cytokines. However, affinity techniques for manufacturing protein therapeutics that use biomolecule-conjugated agarose beads that harness specific biomolecular interactions suffer from issues related to protein denaturation, contamination and the need to maintain biomolecule-specific conditions for efficient protein capture. Here, we report a versatile and scalable method for the purification of recombinant protein therapeutics. The method exploits the high-affinity and controllable host-guest interactions between cucurbit[7]uril (CB[7]) and selected guests such as adamantylammonium. We show that the Herceptin (the brand name of trastuzumab, a monoclonal antibody drug used to treat breast cancer) and the much smaller cytokine interferon α-2a can be purified by site-specifically tagging them with adamantylammonium using the enzyme sortase A, followed by high-affinity binding with CB[7]-conjugated agarose beads and the recovery of the protein using a guest with a stronger affinity for CB[7]. The thermal and chemical stability of CB[7] beads and their scalability, recyclability and low cost may als...Continue Reading

References

Jan 1, 1979·Journal of Immunological Methods·R C DuhamelE Meezan
May 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J C Carrington, W G Dougherty
Mar 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·K WelteM A Moore
Jan 5, 1999·European Journal of Biochemistry·H G SvenssonU Sjöbring
Oct 19, 2000·Methods in Enzymology·J A Bornhorst, J J Falke
Jun 14, 2001·Methods : a Companion to Methods in Enzymology·O PuigB Séraphin
Jun 29, 2002·Journal of Biochemical and Biophysical Methods·Klaus HuseGerhard H Scholz
Apr 2, 2005·Nature Reviews. Drug Discovery·Sven Frokjaer, Daniel E Otzen
Nov 10, 2005·Journal of the American Chemical Society·Simin LiuLyle Isaacs
Oct 7, 2008·Journal of Clinical Pharmacy and Therapeutics·S D Roger, D Goldsmith
Jun 23, 2009·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Andrea HaweWim Jiskoot
Nov 7, 2009·Methods in Enzymology·Helena BlockFrank Schäfer
Nov 10, 2009·Current Opinion in Biotechnology·William R Strohl, David M Knight
Sep 25, 2010·Health Policy·Rodrigo Refoios CamejoRon Herings
Jan 25, 2011·Trends in Biotechnology·Rahul BhambureAnurag S Rathore
Jan 3, 2014·Angewandte Chemie·Liping CaoLyle Isaacs
Feb 19, 2014·The Journal of Physical Chemistry. B·Zsombor Miskolczy, László Biczók
Mar 19, 2014·Trends in Biotechnology·Alexander T Hanke, Marcel Ottens
Oct 16, 2014·Chemical Society Reviews·Khaleel I Assaf, Werner M Nau
Jul 22, 2015·Journal of the American Chemical Society·Mehmet M AyhanOlivier Ouari
Nov 14, 2015·Chemical Reviews·Steven J BarrowOren A Scherman
Dec 6, 2016·Advanced Drug Delivery Reviews·Zehra ElgundiVeysel Kayser
Feb 2, 2017·Angewandte Chemie·James MurrayKimoon Kim
Feb 10, 2017·Journal of the American Chemical Society·David SigwaltLyle Isaacs
Jul 3, 2017·Trends in Pharmacological Sciences·Srinivasa Reddy BonamSylviane Muller
Oct 22, 2019·Journal of the American Chemical Society·Suman Kr GhoshKimoon Kim

❮ Previous
Next ❯

Citations

Jul 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Tian-Tian WangBin Ding
Dec 1, 2020·Journal of the American Chemical Society·Anna Kataki-AnastasakouEllen M Sletten
Sep 7, 2021·Chembiochem : a European Journal of Chemical Biology·Wenbing CaoTao Liu
Nov 4, 2021·Chemical Communications : Chem Comm·Ara LeeKimoon Kim

❮ Previous
Next ❯

Methods Mentioned

BETA
affinity purification
fluorescence imaging
electrophoresis
enzyme-linked immunosorbent assay
flow cytometry

Software Mentioned

Image J

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.