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A comprehensive method to purify three major ANCA antigens: proteinase 3, myeloperoxidase and bactericidal/permeability-increasing protein from human neutrophil granule acid extract.
- Source :
-
Journal of immunological methods [J Immunol Methods] 1996 Oct 16; Vol. 197 (1-2), pp. 121-30. - Publication Year :
- 1996
-
Abstract
- Indirect immunofluorescence (IIF) techniques have shown that anti-neutrophil cytoplasm autoantibodies (ANCA) are useful serological markers for certain small vessel vasculitides and the non-vasculitic inflammatory disorders. ELISA procedures, using purified molecules as solid phase ligands, helped to identify proteinase 3 (PR3) and myeloperoxidase (MPO) as two major ANCA antigens; and recently we characterised bactericidal/permeability-increasing protein (BPI) as another important ANCA antigen. ANCA against these three antigens are associated with different clinical disorders. Therefore purified antigens are needed to determine these different autoantibody specificities in order to help diagnosis and guide treatment. Here we describe a method using Orange-A dye ligand chromatography and cation exchange chromatography for the sequential purification of PR3, MPO and BPI, from the same starting material, an acid extract of normal human neutrophil granules. After separation the three antigens were free of contamination by each other and no traces were found of other known minor ANCA antigens.
- Subjects :
- Antimicrobial Cationic Peptides
Chromatography methods
Humans
Immunoassay methods
Myeloblastin
Neutrophils ultrastructure
Antibodies, Antineutrophil Cytoplasmic immunology
Autoantigens isolation & purification
Blood Proteins isolation & purification
Cytoplasmic Granules chemistry
Membrane Proteins
Neutrophils chemistry
Peroxidase isolation & purification
Serine Endopeptidases isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 0022-1759
- Volume :
- 197
- Issue :
- 1-2
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 8890899
- Full Text :
- https://doi.org/10.1016/0022-1759(96)00123-8