title: Use of anti-(beta2 microglobulin) mAb to study formation of amyloid fibrils.
abstract: Three mAbs, IgG1k 1F11, 7B6 and 14H3, were raised against in vitro-self-aggregated beta2-microglobulin. They recognize the native and unfolded forms of the protein and bind its fibrillar form that is present in amyloid tissue. When assayed in fibrillogenesis tests in vitro, mAb 14H3 inhibited fibril formation from beta2-microglobulin. This mAb recognizes a sequential epitope corresponding to the C-terminal octapeptide, residues 92-99, of beta2-microglobulin. By using synthetic peptides it has been found that the integrity of the sequence is essential for the formation of the immunocomplex: the binding affinity is lowered by one order of magnitude (Kd from 10(-7) M to 10(-6) M) by removal of Met99 and completely abolished when both Asp98 and Met99 are lost or Arg98 is substituted with Lys. The other two mAbs, 1F11 and 7B6, which bind sequences 20-41 and 63-75, respectively, are without effect on beta2-microglobulin fibrillogenesis. These two mAbs recognize beta2-microglobulin bound to the heavy chain in the major histocompatibility complex of type I located in the cell membrane, a property which is not shared by mAb 14H3.
authors.affiliation: Department of Biochemistry, University of Pavia, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Stoppini
authors.forename: V
authors.initials: V
authors.lastname: Bellotti
authors.forename: P
authors.initials: P
authors.lastname: Mangione
authors.forename: G
authors.initials: G
authors.lastname: Merlini
authors.forename: G
authors.initials: G
authors.lastname: Ferri
mesh_terms: D000682:Amyloid / Q000096:biosynthesis* / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D011487:Protein Conformation; D001613:beta 2-Microglobulin / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000682:Amyloid; D000911:Antibodies, Monoclonal; D000939:Epitopes; D001613:beta 2-Microglobulin