Production of conformation-specific monoclonal antibodies against alpha 2 macroglobulin and their use for quantitation of total and transformed alpha 2 macroglobulin in human blood.
Monoclonal antibodies against the human proteinase inhibitor, alpha 2 macroglobulin, have been produced by immunizing BALB/c mice with alpha 2 macroglobulin reacted with methylamine. Two antibodies have been characterized in detail with respect to their binding to native alpha 2 macroglobulin and to different derivatives of the inhibitor. The antibody alpha-1 was found to recognize only those forms of the inhibitor which were transformed by reaction with different proteinases or with methylamine. Binding of alpha-1 was mapped to a specific epitope localized within a distance of 138 amino acid residues from the C terminal end of alpha 2 macroglobulin. The C terminal end is assumed to be exposed during the transformation of the inhibitor and harbours the receptor recognition site. The monoclonal antibody alpha-11 was found to bind to all forms of the inhibitor indicating that its epitope is located in a region not involved in major conformational changes of the inhibitor. On the basis of the different reactivity patterns of alpha-1 and alpha-11 two enzyme-linked immunosorption assays were established for quantitation of total and transformed alpha 2 macroglobulin in human blood. The concentration of the two forms have been determined in a population of 114 healthy individuals giving values of 254 +/- 6.6 mg/dl (mean +/- SEM) of total alpha 2 macroglobulin and 1.07 +/- 0.05 mg/dl (mean +/- SEM) of the transformed inhibitor.
Birkenmeier
G
G
Institute of Biochemistry, University of Leipzig, Germany.
Stigbrand
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000917:Antibody Formation; D001666:Binding Sites, Antibody; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000737:chemistry / Q000378:metabolism; D005260:Female; D006801:Humans; D007527:Isoenzymes / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D011487:Protein Conformation; D000511:alpha-Macroglobulins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007527:Isoenzymes; D000511:alpha-Macroglobulins