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Generation and Characterization of Native and Sialic Acid-Deficient IgE.
Alex J McCraw1, Richard A Gardner2, Anna M Davies3
1St. John's Institute of Dermatology, School of Basic & Medical Biosciences, King's College London, London SE1 9RT, UK.
International Journal of Molecular Sciences
|November 11, 2022
Summary
Researchers generated and analyzed native and sialic acid-deficient human IgE antibodies. These glyco-engineered antibodies maintain functional competence, crucial for allergy and cancer immunotherapy research.
Area of Science:
- Immunology
- Glycobiology
- Biotechnology
Background:
- Characterizing IgE antibodies and their glycans is vital for allergy and AllergoOncology.
- Understanding IgE glycosylation impacts antibody function in immunotherapy.
Purpose of the Study:
- To generate, glyco-profile, and functionally analyze native and sialic acid-deficient human IgE.
- To assess the impact of sialic acid deficiency on IgE structure and function.
Main Methods:
- Production and purification of human IgE using affinity chromatography.
- Glycan analysis via Hydrophilic interaction liquid chromatography (HILIC)-high performance liquid chromatography (HPLC).
- Functional assays including Fc receptor binding and degranulation studies.
Main Results:
- Successfully generated and purified native and sialic acid-deficient IgE antibodies.
- Glyco-profiling confirmed sialic acid loss and altered glycan profiles in engineered IgE.
- Both native and engineered IgE demonstrated Fc receptor recognition and triggered cell degranulation.
Conclusions:
- Efficient generation of functional recombinant native and sialic acid-deficient IgE is demonstrated.
- Glyco-engineering of IgE is feasible and maintains antibody functionality.
- These findings support the use of engineered IgE in allergy and AllergoOncology.
