{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T09:54:20Z","timestamp":1764842060536,"version":"build-2065373602"},"reference-count":28,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2018,4,16]],"date-time":"2018-04-16T00:00:00Z","timestamp":1523836800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","award":["01GM1516C"],"award-info":[{"award-number":["01GM1516C"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemMedChem"],"published-print":{"date-parts":[[2018,5,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>\u03b1\u2010Galactosidase (\u03b1Gal) is a lysosomal enzyme that hydrolyses the terminal \u03b1\u2010galactosyl moiety from glycosphingolipids. Mutations in the encoding genes for \u03b1Gal lead to defective or misfolded enzyme, which results in substrate accumulation and subsequent organ dysfunction. The metabolic disease caused by a deficiency of human \u03b1\u2010galactosidase\u2005A is known as Fabry disease or Fabry\u2013Anderson disease, and it belongs to a larger group known as lysosomal storage diseases. An effective treatment for Fabry disease has been developed by enzyme replacement therapy (ERT), which involves infusions of purified recombinant enzyme in order to increase enzyme levels and decrease the amounts of accumulated substrate. However, immunoreactivity and IgG antibody formation are major, therapy\u2010limiting, and eventually life\u2010threatening complications of ERT. The present study focused on the epitope determination of human \u03b1\u2010galactosidase\u2005A against its antibody formed. Here we report the identification of the epitope of human \u03b1Gal(309\u2013332) recognized by a human monoclonal anti\u2010\u03b1Gal antibody, using a combination of proteolytic excision of the immobilized immune complex and surface plasmon resonance biosensing mass spectrometry. The epitope peptide, \u03b1Gal(309\u2013332), was synthesized by solid\u2010phase peptide synthesis. Determination of its affinity by surface plasmon resonance analysis revealed a high binding affinity for the antibody (<jats:italic>K<\/jats:italic><jats:sub>D<\/jats:sub>=39\u00d710<jats:sup>\u22129<\/jats:sup>\u2009<jats:sc>m<\/jats:sc>), which is nearly identical to that of the full\u2010length enzyme (<jats:italic>K<\/jats:italic><jats:sub>D<\/jats:sub>=16\u00d710<jats:sup>\u22129<\/jats:sup>\u2009<jats:sc>m<\/jats:sc>). The proteolytic excision affinity mass spectrometry method is shown here to be an efficient tool for epitope identification of an immunogenic lysosomal enzyme. Because the full\u2010length \u03b1Gal and the antibody epitope showed similar binding affinities, this provides a basis for reversing immunogenicity upon ERT by: 1)\u2005treatment of patients with the epitope peptide to neutralize antibodies, or 2)\u2005removal of antibodies by apheresis, and thus significantly improving the response to ERT.<\/jats:p>","DOI":"10.1002\/cmdc.201800094","type":"journal-article","created":{"date-parts":[[2018,2,23]],"date-time":"2018-02-23T05:26:39Z","timestamp":1519363599000},"page":"909-915","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Antibody Epitope of Human \u03b1\u2010Galactosidase\u2005A Revealed by Affinity Mass Spectrometry: A Basis for Reversing Immunoreactivity in Enzyme Replacement Therapy of Fabry Disease"],"prefix":"10.1002","volume":"13","author":[{"given":"Zdenek","family":"Kukacka","sequence":"first","affiliation":[{"name":"Steinbeis Centre for Biopolymer Analysis and Biomedical Mass Spectrometry  65428 R\u00fcsselsheim am Main Germany"},{"name":"Department of Chemistry University of Konstanz  78457 Konstanz Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marius","family":"Iurascu","sequence":"additional","affiliation":[{"name":"Steinbeis Centre for Biopolymer Analysis and Biomedical Mass Spectrometry  65428 R\u00fcsselsheim am Main Germany"},{"name":"Department of Chemistry University of Konstanz  78457 Konstanz Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Loredana","family":"Lupu","sequence":"additional","affiliation":[{"name":"Steinbeis Centre for Biopolymer Analysis and Biomedical Mass Spectrometry  65428 R\u00fcsselsheim am Main Germany"},{"name":"Department of Chemistry University of Konstanz  78457 Konstanz Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hendrik","family":"Rusche","sequence":"additional","affiliation":[{"name":"Steinbeis Centre for Biopolymer Analysis and Biomedical Mass Spectrometry  65428 R\u00fcsselsheim am Main Germany"},{"name":"Department of Chemistry University of Konstanz  78457 Konstanz Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mary","family":"Murphy","sequence":"additional","affiliation":[{"name":"Ametek Reichert Technologies  Depew NY 14043 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lorenzo","family":"Altamore","sequence":"additional","affiliation":[{"name":"French\u2013Italian Interdepartmental Laboratory of Peptide and Protein Chemistry and Biology Universit\u00e0 degli Studi di Firenze  50019 Sesto Fiorentino Italy"},{"name":"Dipartimento di Chimica \u201cUgo Schiff\u201d Universit\u00e0 degli Studi di Firenze  Via della Lastruccia 13 50019 Sesto Fiorentino Italy"},{"name":"PeptLab@UCP and Laboratory of Chemical Biology EA4505 Universit\u00e9 Paris-Seine  5 Mail Gay-Lussac 95031 Cergy-Pontoise France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fabio","family":"Borri","sequence":"additional","affiliation":[{"name":"French\u2013Italian Interdepartmental Laboratory of Peptide and Protein Chemistry and Biology Universit\u00e0 degli Studi di Firenze  50019 Sesto Fiorentino Italy"},{"name":"Dipartimento di Chimica \u201cUgo Schiff\u201d Universit\u00e0 degli Studi di Firenze  Via della Lastruccia 13 50019 Sesto Fiorentino Italy"},{"name":"PeptLab@UCP and Laboratory of Chemical Biology EA4505 Universit\u00e9 Paris-Seine  5 Mail Gay-Lussac 95031 Cergy-Pontoise France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefan","family":"Maeser","sequence":"additional","affiliation":[{"name":"Steinbeis Centre for Biopolymer Analysis and Biomedical Mass Spectrometry  65428 R\u00fcsselsheim am Main Germany"},{"name":"Department of Chemistry University of Konstanz  78457 Konstanz Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna Maria","family":"Papini","sequence":"additional","affiliation":[{"name":"French\u2013Italian Interdepartmental Laboratory of Peptide and Protein Chemistry and Biology Universit\u00e0 degli Studi di Firenze  50019 Sesto Fiorentino Italy"},{"name":"Dipartimento di Chimica \u201cUgo Schiff\u201d Universit\u00e0 degli Studi di Firenze  Via della Lastruccia 13 50019 Sesto Fiorentino Italy"},{"name":"PeptLab@UCP and Laboratory of Chemical Biology EA4505 Universit\u00e9 Paris-Seine  5 Mail Gay-Lussac 95031 Cergy-Pontoise France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julia","family":"Hennermann","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Villa Metabolica Universit\u00e4tsmedizin Mainz  55130 Mainz Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2611-5389","authenticated-orcid":false,"given":"Michael","family":"Przybylski","sequence":"additional","affiliation":[{"name":"Steinbeis Centre for Biopolymer Analysis and Biomedical Mass Spectrometry  65428 R\u00fcsselsheim am Main Germany"},{"name":"Department of Chemistry University of Konstanz  78457 Konstanz 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