{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T16:36:40Z","timestamp":1780591000616,"version":"3.54.1"},"reference-count":33,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,2,21]],"date-time":"2021-02-21T00:00:00Z","timestamp":1613865600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The C-terminal-fragments of alpha1-antitrypsin (AAT) have been identified and their diverse biological roles have been reported in vitro and in vivo. These findings prompted us to develop a monoclonal antibody that specifically recognizes C-36 peptide (corresponding to residues 359\u2013394) resulting from the protease-associated cleavage of AAT. The C-36-targeting mouse monoclonal Immunoglobulin M (IgM) antibody (containing \u03ba light chains, clone C42) was generated and enzyme-linked immunosorbent assay (ELISA)-tested by Davids Biotechnologie GmbH, Germany. Here, we addressed the effectiveness of the novel C42 antibody in different immunoassay formats, such as dot- and Western blotting, confocal laser microscopy, and flow cytometry. According to the dot-blot results, our novel C42 antibody detects the C-36 peptide at a range of 0.1\u20130.05 \u00b5g and shows no cross-reactivity with native, polymerized, or oxidized forms of full-length AAT, the AAT-elastase complex mixture, as well as with shorter C-terminal fragments of AAT. However, the C42 antibody does not detect denatured peptide in SDS-PAGE\/Western blotting assays. On the other hand, our C42 antibody, unconjugated as well as conjugated to DyLight488 fluorophore, when applied for immunofluorescence microscopy and flow cytometry assays, specifically detected the C-36 peptide in human blood cells. Altogether, we demonstrate that our novel C42 antibody successfully recognizes the C-36 peptide of AAT in a number of immunoassays and has potential to become an important tool in AAT-related studies.<\/jats:p>","DOI":"10.3390\/ijms22042141","type":"journal-article","created":{"date-parts":[[2021,2,21]],"date-time":"2021-02-21T22:04:15Z","timestamp":1613945055000},"page":"2141","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["A Novel Mouse Monoclonal Antibody C42 against C-Terminal Peptide of Alpha-1-Antitrypsin"],"prefix":"10.3390","volume":"22","author":[{"given":"Srinu","family":"Tumpara","sequence":"first","affiliation":[{"name":"Department of Respiratory Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6327-6862","authenticated-orcid":false,"given":"Elena","family":"Korenbaum","sequence":"additional","affiliation":[{"name":"Institute for Biophysical Chemistry, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark","family":"K\u00fchnel","sequence":"additional","affiliation":[{"name":"Institute of Pathology, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danny","family":"Jonigk","sequence":"additional","affiliation":[{"name":"Institute of Pathology, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beata","family":"Olejnicka","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine, Lund University, SE-221 00 Lund, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Davids","sequence":"additional","affiliation":[{"name":"Davids Biotechnologie GmbH, R\u00f6ntgenstra\u00dfe 3, 93055 Regensburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tobias","family":"Welte","sequence":"additional","affiliation":[{"name":"Department of Respiratory Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6834-350X","authenticated-orcid":false,"given":"Beatriz","family":"Martinez-Delgado","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics, Institute of Health Carlos III, Center for Biomedical Research in the Network of Rare Diseases (CIBERER), 28220 Majadahonda, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabina","family":"Janciauskiene","sequence":"additional","affiliation":[{"name":"Department of Respiratory Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), Hannover Medical School, 30625 Hannover, Germany"},{"name":"Department of Genetics and Clinical Immunology, National Institute of Tuberculosis and Lung Diseases, 01-138 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,2,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/S0925-4439(01)00025-4","article-title":"Conformational Properties of Serine Proteinase Inhibitors (Serpins) Confer Multiple Pathophysiological Roles","volume":"1535","author":"Janciauskiene","year":"2001","journal-title":"Biochim Biophys Acta"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1016\/S0934-8832(11)80066-1","article-title":"Isolation and Serine Protease Inhibitory Activity of the 44-Residue, C-Terminal Fragment of Alpha 1-Antitrypsin from Human Placenta","volume":"12","author":"Niemann","year":"1992","journal-title":"Matrix"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1007\/s10549-009-0625-5","article-title":"A Unique Proteolytic Fragment of Alpha1-Antitrypsin Is Elevated in Ductal Fluid of Breast Cancer Patient","volume":"123","author":"Zhou","year":"2010","journal-title":"Breast Cancer Res. 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