title: Epitope recognition in the human-pig comparison model on fixed and embedded material.
abstract: The conditions and the specificity by which an antibody binds to its target protein in routinely fixed and embedded tissues are unknown. Direct methods, such as staining in a knock-out animal or in vitro peptide scanning of the epitope, are costly and impractical. We aimed to elucidate antibody specificity and binding conditions using tissue staining and public genomic and immunological databases by comparing human and pig-the farmed mammal evolutionarily closest to humans besides apes. We used a database of 146 anti-human antibodies and found that antibodies tolerate partially conserved amino acid substitutions but not changes in target accessibility, as defined by epitope prediction algorithms. Some epitopes are sensitive to fixation and embedding in a species-specific fashion. We also find that half of the antibodies stain porcine tissue epitopes that have 60% to 100% similarity to human tissue at the amino acid sequence level. The reason why the remaining antibodies fail to stain the tissues remains elusive. Because of its similarity with the human, pig tissue offers a convenient tissue for quality control in immunohistochemistry, within and across laboratories, and an interesting model to investigate antibody specificity.
authors.affiliation: Azienda Ospedaliera San Gerardo, Monza, Italy (CRS, RG, LR, LT, AM, GC, MMB)
authors.forename: Carla Rossana
authors.initials: CR
authors.lastname: Scalia
authors.affiliation: Azienda Ospedaliera San Gerardo, Monza, Italy (CRS, RG, LR, LT, AM, GC, MMB)
authors.forename: Rossella
authors.initials: R
authors.lastname: Gendusa
authors.affiliation: Dipartimento di Chirurgia e Medicina Traslazionale, Universitá degli Studi di Milano-Bicocca, Monza Italy (MB, GC)
authors.forename: Maria
authors.initials: M
authors.lastname: Basciu
authors.affiliation: Azienda Ospedaliera San Gerardo, Monza, Italy (CRS, RG, LR, LT, AM, GC, MMB)
authors.forename: Lorella
authors.initials: L
authors.lastname: Riva
authors.affiliation: Azienda Ospedaliera San Gerardo, Monza, Italy (CRS, RG, LR, LT, AM, GC, MMB)
authors.forename: Lorenza
authors.initials: L
authors.lastname: Tusa
authors.affiliation: Azienda Ospedaliera San Gerardo, Monza, Italy (CRS, RG, LR, LT, AM, GC, MMB)
authors.forename: Antonella
authors.initials: A
authors.lastname: Musarò
authors.affiliation: Struttura Complessa di Anatomia Patologica, Dipartimento di Medicina di Laboratorio, Azienda Ospedaliera Ospedale Niguarda Ca' Granda, Milano Italy (SV)
authors.forename: Silvio
authors.initials: S
authors.lastname: Veronese
authors.affiliation: Servizio di Igiene degli Alimenti di Origine Animale, Dipartimento Veterinario, Azienda Sanitaria Locale di Monza e Brianza, Desio, Italy (AF, DD)
authors.forename: Angelo
authors.initials: A
authors.lastname: Formenti
authors.affiliation: Servizio di Igiene degli Alimenti di Origine Animale, Dipartimento Veterinario, Azienda Sanitaria Locale di Monza e Brianza, Desio, Italy (AF, DD)
authors.forename: Donatella
authors.initials: D
authors.lastname: D'Angelo
authors.affiliation: Dipartimento di Prevenzione Veterinario, Distretto Veterinario 2 Legnano - Castano Primo, Azienda Sanitaria Locale Milano 1, Castano Primo, Italy (AGR)
authors.forename: Angela Gabriella
authors.initials: AG
authors.lastname: Ronzio
authors.affiliation: Azienda Ospedaliera San Gerardo, Monza, Italy (CRS, RG, LR, LT, AM, GC, MMB)|Dipartimento di Chirurgia e Medicina Traslazionale, Universitá degli Studi di Milano-Bicocca, Monza Italy (MB, GC)
authors.forename: Giorgio
authors.initials: G
authors.lastname: Cattoretti
authors.affiliation: Azienda Ospedaliera San Gerardo, Monza, Italy (CRS, RG, LR, LT, AM, GC, MMB)
authors.forename: Maddalena Maria
authors.initials: MM
authors.lastname: Bolognesi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology; D000918:Antibody Specificity; D018604:Epitope Mapping / Q000379:methods; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008969:Molecular Sequence Data; D016612:Paraffin Embedding / Q000379:methods; D013552:Swine; D016707:Tissue Fixation / Q000379:methods
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes
keywords: antigen; epitope; immunohistochemistry; quality control; swine