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In this study, we employed an imaging mass spectrometry\u2010based workflow called MALDI\u2010IHC, which uses antibodies conjugated with photocleavable mass\u2010tags, to investigate the detection of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) along with pan\u2010cytokeratin in formalin\u2010fixed paraffin\u2010embedded (FFPE) sections from cases with breast cancer. The expression of ER, PR, and HER2 was consistent with the clinical diagnosis based on conventional immunohistochemistry, lacking signal in triple\u2010negative cases and concordant signals in the cases that were classified as ER+\/PR+\/HER2+, ER+\/PR+\/HER2\u2212, and ER\u2212\/PR\u2212\/HER2+ groups, respectively. In addition, the potential utility of a seconday antibody\u2010based MALDI\u2010IHC approach for HER2 expression was evaluated. In agreement with immunohistochemistry results, HER2 expression co\u2010localized within tumor regions in a HER2+ breast cancer case, while no expression was observed in a HER2\u2212 case. Collectively, these results highlight the potential of imaging mass spectrometry\u2010based MALDI\u2010IHC workflow for multiplexed detection of proteins in clinically relevant tissue sections.<\/jats:p>","DOI":"10.1002\/pmic.70110","type":"journal-article","created":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T21:25:47Z","timestamp":1773005147000},"page":"6-12","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Imaging Mass Spectrometry\u2010Based Assessment of ER, PR, and HER2 Protein Expression in Breast Cancer"],"prefix":"10.1002","volume":"26","author":[{"given":"Kiran K.","family":"Mangalaparthi","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daigo","family":"Gunji","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olajide E.","family":"Olaleye","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amy J.","family":"French","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gunveen S.","family":"Sachdeva","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"},{"name":"Manipal Academy of Higher Education  Manipal Karnataka India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shilpa","family":"Venkataraman","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"},{"name":"Manipal Academy of Higher Education  Manipal Karnataka India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Neha","family":"Joshi","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tianqi","family":"Gao","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sumankalai","family":"Ramachandran","sequence":"additional","affiliation":[{"name":"Bruker Scientific LLC  Billerica Massachusetts USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julie A.","family":"Vrana","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kenneth J.","family":"Rothschild","sequence":"additional","affiliation":[{"name":"AmberGen Inc.  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