Monoclonal antibodies to rabbit alpha-2-macroglobulin and their use in a sensitive ELISA assay.
alpha-2-macroglobulin (alpha-2-M), a plasma proteinase inhibitor, plays an important role in the pathogenesis of lung inflammation. The purpose of this study was to develop a sensitive ELISA assay for rabbit alpha-2-M to allow us to define the role of this protein in a rabbit model of lung inflammation/injury. Therefore, we developed hybridomas which secrete monoclonal antibodies (mAbs) against rabbit alpha-2-M. From the antibodies produced, two (5B6 and 3C5) were selected. Both of them were of the IgG1 subclass. 5B6 reacted with native alpha-2-M as well as with the "fast" form of alpha-2-M (native alpha-2-M or the "slow" form is converted to the "fast" form by reaction with a proteinase). On the other hand, 3C5 reacted only with "fast" form of alpha-2-M. With these antibodies, we developed two ELISA assays which were used to determine the concentration of alpha-2-M in the lung fluids from rabbits with lung injury.
Kurdowska
Anna
A
Department of Biochemistry, University of Texas Health Center at Tyler, 11937 US Highway 271, Tyler, TX 75708-3154, USA. anna.kudowska@uthct.edu
Miller
Edmund J
EJ
Krupa
Agnieszka
A
Noble
James M
JM
Sakao
Yukinori
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D001992:Bronchoalveolar Lavage Fluid; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D011817:Rabbits; D012680:Sensitivity and Specificity; D000511:alpha-Macroglobulins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000511:alpha-Macroglobulins