Generation and characterization of monoclonal antibodies against the intracellular domain of hemidesmosomal type XVII collagen.
Type XVII collagen, also referred to as bullous pemphigoid antigen 2 (BPAG2) or bullous pemphigoid antigen 180 (BP180), is a transmembrane protein of the hemidesmosomal complexes of keratinocytes. Type XVII collagen has an unusual type II orientation with its N-terminus intracellularly located and with a large extracellular domain that spans lamina lucida of the dermal-epidermal junction. Type XVII collagen is an autoantigen in patients with pemphigoid diseases and its gene is mutated in patients with junctional epidermolysis bullosa. In the present work, we generated new monoclonal antibodies (MAbs) against the intracellular domain of type XVII collagen. We further characterized reactivity and fine specificity of an MAb (clone V58) from this panel of antibodies. The epitope recognized by the mAb V58 was mapped to a stretch of type XVII collagen corresponding to residues 234-398 of its sequence. Possible applications of this new MAb include antigen mapping in patients with hereditary epidermolysis bullosa and immunoaffinity purification of cell-derived type XVII collagen.
Olaru
Florina
F
Department of Dermatology, University of Lübeck, Lübeck, Romania.
Mihai
Sidonia
S
Petrescu
Ioan
I
Zillikens
Detlef
D
Sitaru
Cassian
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology*; D002352:Carrier Proteins; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D000939:Epitopes; D005260:Female; D022002:Hemidesmosomes / Q000276:immunology*; D006801:Humans; D007424:Intracellular Fluid / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens / Q000737:chemistry / Q000276:immunology*; D017434:Protein Structure, Tertiary; D000097805:Collagen Type XVII
D000911:Antibodies, Monoclonal; D001324:Autoantigens; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D000939:Epitopes; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D000097805:Collagen Type XVII; C486268:DST protein, human