title: Validation and comparison of anti-αvβ3 and anti-αvβ5 rabbit monoclonal versus murine monoclonal antibodies in four different tumor entities.
abstract: Integrins are pivotal in cancer biology and are putative candidates for cancer therapy. The investigation of integrins has been hampered by the lack of antibodies suitable for formalin-fixed and paraffin-embedded (FFPE) tissue specimens. Here, we validated monoclonal rabbit antibodies (RabMAbs) against integrins αvβ3 (EM22703) and αvβ5 (EM09902) with murine monoclonal antibodies (MuMabs) LM609 (against αvβ3) and P1F6 (against αvβ5), in immunohistochemistry. Immunostaining was performed on sections of matching unfixed, cryoconserved (CC) and FFPE tissue from 19 colorectal, 20 lung, 17 breast, and 9 ovarian carcinomas. Sections were stained with LM609 and P1F6 and compared with the immunoreactions of the RabMAbs. The degree of concordance was assigned for staining patterns and intensity. Concordance between MuMAbs and RabMAbs ranged from weak, for anti-αvβ5 antibodies, to nearly complete for anti-αvβ3 antibodies. We confirmed that MuMAbs LM609 and P1F6 bound very weakly in FFPE tissue and no staining was seen. By contrast, RabMAbs EM22703 and EM09902 generally showed a high degree of agreement in staining patterns of CC and FFPE tissue. In summary, the RabMAbs had overlapping staining patterns that were generally more intense for CC when compared with FFPE material. This study suggests that EM22703 and EM09902 staining closely matches the staining of standard MuMAbs and also does so in archival FFPE tissue.
authors: Departments of *Pathology †Experimental Cancer Research, Christian-Albrechts-University, Kiel ‡Department of Biochemistry and Cellular Pharmacology, Oncology Platform, Merck KGaA, Darmstadt, Germany.
authors: Christine
authors: C
authors: Böger
authors: Holger
authors: H
authors: Kalthoff
authors: Simon L
authors: SL
authors: Goodman
authors: Christoph
authors: C
authors: Röcken
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D002277:Carcinoma / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D015179:Colorectal Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D005260:Female; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D039302:Integrin alphaVbeta3 / Q000032:analysis* / Q000235:genetics; D008175:Lung Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D008297:Male; D051379:Mice; D008875:Middle Aged; D009369:Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D010051:Ovarian Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D016612:Paraffin Embedding; D011817:Rabbits; D019030:Receptors, Vitronectin / Q000032:analysis* / Q000235:genetics; D013045:Species Specificity; D016707:Tissue Fixation
chemical_list: D000911:Antibodies, Monoclonal; D039302:Integrin alphaVbeta3; D019030:Receptors, Vitronectin; C064660:integrin alphaVbeta5