Transient pentameric IgM fulfill biological function-Effect of expression host and transfection on IgM properties

Recombinant production of IgM antibodies poses a special challenge due to the complex structure of the proteins and their not yet fully elucidated interactions with the immune effector proteins, especially the complement system. In this study, we present transient expression of IgM antibodies (IgM61...

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Bibliographic Details
Published in:PloS one 2020-03, Vol.15 (3), p.e0229992-e0229992
Main Authors: Hennicke, Julia, Schwaigerlehner, Linda, Grünwald-Gruber, Clemens, Bally, Isabelle, Ling, Wai Li, Thielens, Nicole, Reiser, Jean-Baptiste, Kunert, Renate
Format: Article
Language:English
Subjects:
Antibodies
Antigens
Biochemistry, Molecular Biology
Biological effects
Biology and Life Sciences
Biotechnology
Comparative analysis
Complement
Complement activation
Cytomegalovirus
Dihydrofolate reductase
Electron microscopy
Genes
Glycan
Glycosylation
Immunoglobulin M
Life Sciences
Mediation
Medicine and Health Sciences
Microscopy
Natural resources
Physical Sciences
Polymers
Post-translation
Production management
Proteins
Quality management
Research and analysis methods
Structural Biology
Structural integrity
Structure-function relationships
Transfection
Transmission electron microscopy
Workflow
Workflow software
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Summary:Recombinant production of IgM antibodies poses a special challenge due to the complex structure of the proteins and their not yet fully elucidated interactions with the immune effector proteins, especially the complement system. In this study, we present transient expression of IgM antibodies (IgM617, IgM012 and IgM012_GL) in HEK cells and compared it to the well-established stable expression system in CHO cells. The presented workflow investigates quality attributes including productivity, polymer distribution, glycosylation, antibody structure and activation of the classical complement pathway. The HEK293E transient expression system is able to generate comparable amounts and polymer distribution as IgM stably produced in CHO. Although the glycan profile generated by HEK293E cells contained a lower degree of sialylation and a higher portion of oligomannose structures, the potency to activate the complement cascade was maintained. Electron microscopy also confirmed the structural integrity of IgM pentamers produced in HEK293E cells, since the conventional star-shaped structure is observed. From our studies, we conclude that the transient expression system provides an attractive alternative for rapid, efficient and high-throughput production of complex IgM antibodies with slightly altered post-translational modifications, but comparable structure and function.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0229992