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We report the generation, glyco-profiling and functional analysis of native and sialic acid-deficient glyco-engineered human IgE. The antibodies produced from human embryonic kidney cells were purified via a human IgE class-specific affinity matrix and structural integrity was confirmed by SDS-PAGE and size-exclusion chromatography (SEC). Purified IgEs specific for the tumor-associated antigens Chondroitin Sulfate Proteoglycan 4 (CSPG4-IgE) and Human Epidermal Growth Factor Receptor 2 (HER2-IgE) were devoid of by-products such as free light chains. Using neuraminidase-A, we generated sialic acid-deficient CSPG4-IgE as example glyco-engineered antibody. Comparative glycan analyses of native and glyco-engineered IgEs by Hydrophilic interaction liquid chromatography (HILIC)-high performance liquid chromatography (HPLC) indicated loss of sialic acid terminal residues and differential glycan profiles. Native and glyco-engineered CSPG4-IgEs recognized Fc receptors on the surface of human Fc\u03b5RI-expressing rat basophilic leukemia RBL-SX38 cells, and of CD23\/Fc\u03b5RII-expressing human RPMI-8866 B-lymphocytes and bound to CSPG4-expressing A2058 human melanoma cells, confirming Fab-mediated recognition. When cross-linked on the cell surface, both IgEs triggered RBL-SX38 degranulation. We demonstrate efficient generation and functional competence of recombinant native and sialic acid-deficient IgEs.<\/jats:p>","DOI":"10.3390\/ijms232113455","type":"journal-article","created":{"date-parts":[[2022,11,4]],"date-time":"2022-11-04T03:28:10Z","timestamp":1667532490000},"page":"13455","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Generation and Characterization of Native and Sialic Acid-Deficient IgE"],"prefix":"10.3390","volume":"23","author":[{"given":"Alex J.","family":"McCraw","sequence":"first","affiliation":[{"name":"St. John\u2019s Institute of Dermatology, School of Basic & Medical Biosciences, King\u2019s College London, London SE1 9RT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7601-7544","authenticated-orcid":false,"given":"Richard A.","family":"Gardner","sequence":"additional","affiliation":[{"name":"Ludger, Ltd., Culham Science Centre, Abingdon, Oxfordshire OX14 3EB, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1498-1594","authenticated-orcid":false,"given":"Anna M.","family":"Davies","sequence":"additional","affiliation":[{"name":"Randall Centre for Cell and Molecular Biophysics, School of Basic & Medical Biosciences, King\u2019s College London, London SE1 9RT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6386-0890","authenticated-orcid":false,"given":"Daniel I. R.","family":"Spencer","sequence":"additional","affiliation":[{"name":"Ludger, Ltd., Culham Science Centre, Abingdon, Oxfordshire OX14 3EB, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Melanie","family":"Grandits","sequence":"additional","affiliation":[{"name":"St. John\u2019s Institute of Dermatology, School of Basic & Medical Biosciences, King\u2019s College London, London SE1 9RT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gerd K.","family":"Wagner","sequence":"additional","affiliation":[{"name":"St. John\u2019s Institute of Dermatology, School of Basic & Medical Biosciences, King\u2019s College London, London SE1 9RT, UK"},{"name":"Medical Biology Centre, School of Pharmacy, Queen\u2019s University Belfast, 97 Lisburn Road, Belfast BT9 7BL, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James M.","family":"McDonnell","sequence":"additional","affiliation":[{"name":"Randall Centre for Cell and Molecular Biophysics, School of Basic & Medical Biosciences, King\u2019s College London, London SE1 9RT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4100-7810","authenticated-orcid":false,"given":"Sophia N.","family":"Karagiannis","sequence":"additional","affiliation":[{"name":"St. John\u2019s Institute of Dermatology, School of Basic & Medical Biosciences, King\u2019s College London, London SE1 9RT, UK"},{"name":"Breast Cancer Now Research Unit, School of Cancer & Pharmaceutical Sciences, Guy\u2019s Cancer Centre, King\u2019s College London, London SE1 9RT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alicia","family":"Chenoweth","sequence":"additional","affiliation":[{"name":"St. John\u2019s Institute of Dermatology, School of Basic & Medical Biosciences, King\u2019s College London, London SE1 9RT, UK"},{"name":"Breast Cancer Now Research Unit, School of Cancer & Pharmaceutical Sciences, Guy\u2019s Cancer Centre, King\u2019s College London, London SE1 9RT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1909-5957","authenticated-orcid":false,"given":"Silvia","family":"Crescioli","sequence":"additional","affiliation":[{"name":"St. John\u2019s Institute of Dermatology, School of Basic & Medical Biosciences, King\u2019s College London, London SE1 9RT, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1146\/annurev.immunol.21.120601.141103","article-title":"The biology of IGE and the basis of allergic disease","volume":"21","author":"Gould","year":"2003","journal-title":"Annu. 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