Site-specific immobilization of endoglycosidases for streamlined chemoenzymatic glycan remodeling of antibodies

Carbohydrate Research
Tiezheng LiLai-Xi Wang

Abstract

Chemoenzymatic glycan remodeling of antibodies using an endoglycosidase and its mutant is emerging as an attractive approach for producing homogeneous antibody glycoforms. We report in this paper a site-specific covalent immobilization of the endoglycosidases (Endo-S2 and its glycosynthase mutant D184M) using a recombinant microbial transglutaminase (MTG) and evaluation of the immobilized enzymes in deglycosylation and glycosylation of a therapeutic antibody. The site-specific covalent immobilization was achieved by introduction of a Q-tag at the C-terminus of the recombinant enzymes followed by conjugation of the enzymes to a primary amine-containing solid support through MTG-catalyzed transglutamination. Using rituximab as a model system, we found that the Endo-S2 wild-type and D184M glycosynthase mutant immobilized by this approach were efficient in the two step antibody glycan remodeling to generate homogeneous antibody glycoforms. Notably using the covalently immobilized enzymes can efficiently avoid the need of intermediate purification and eliminate the residual contamination of wild type enzyme for product hydrolysis, thus streamlining the chemoenzymatic Fc glycan remodeling of antibodies.

References

Jan 1, 1992·Analytical Biochemistry·M BondeD S Pepper
Sep 28, 2005·Biotechnology and Bioengineering·Dale WarnockRobert J Bayer
Dec 3, 2005·Science·Falk Nimmerjahn, Jeffrey V Ravetch
Aug 5, 2006·Science·Yoshikatsu KanekoJeffrey V Ravetch
Jun 23, 2007·Journal of Biomedical Materials Research. Part a·C W YungT A Barbari
Apr 19, 2008·Science·Robert M AnthonyJeffrey V Ravetch
Mar 5, 2011·The International Journal of Artificial Organs·Li-Qun WuGregory F Payne
Jul 20, 2011·Proceedings of the National Academy of Sciences of the United States of America·Claudia FerraraJörg Benz
Jul 4, 2012·Journal of the American Chemical Society·Wei HuangLai-Xi Wang
Apr 26, 2013·Applied Microbiology and Biotechnology·Maria T GundersenJoelle N Pelletier
Jan 21, 2014·Chemistry & Biology·Lai-Xi Wang, Mohammed N Amin
May 16, 2014·Chemical Communications : Chem Comm·Samuel K Oteng-PabiJeffrey W Keillor
Mar 4, 2015·Proceedings of the National Academy of Sciences of the United States of America·Nathaniel WashburnCarlos J Bosques
Aug 9, 2015·Proceedings of the National Academy of Sciences of the United States of America·Chin-Wei LinChi-Huey Wong
Oct 27, 2015·ACS Biomaterials Science & Engineering·Yi LiuGregory F Payne
Oct 30, 2016·Metabolic Engineering·Narendranath BhokishamWilliam E Bentley
Aug 2, 2016·Biotechnology and Bioengineering·Hsuan-Chen WuWilliam E Bentley
Mar 16, 2017·Proceedings of the National Academy of Sciences of the United States of America·Tiezheng LiLai-Xi Wang
May 12, 2017·Carbohydrate Research·Weilai GuoWei Huang

❮ Previous
Next ❯

Citations

Oct 7, 2018·Biological Chemistry·Lukas DeweidHarald Kolmar
Dec 28, 2019·World Journal of Microbiology & Biotechnology·Lovaine DuarteMarco Antônio Záchia Ayub
Mar 28, 2019·Annual Review of Biochemistry·Lai-Xi WangTiezheng Li
Mar 31, 2021·Biotechnology Advances·Zuzana MészárosKristýna Slámová
Apr 23, 2021·Chemical Record : an Official Publication of the Chemical Society of Japan ... [et Al.]·Shino Manabe, Yoshiki Yamaguchi
Aug 25, 2018·Chemical Reviews·Chao Li, Lai-Xi Wang
Feb 5, 2019·Journal of the American Chemical Society·Larissa Krasnova, Chi-Huey Wong
Sep 1, 2021·Biotechnology Advances·Zuzana MészárosKristýna Slámová

❮ Previous
Next ❯

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.