Immunochemistry of human Lp[a]: characterization of monoclonal antibodies that cross-react strongly with plasminogen.
Forty different monoclonal antibodies were produced from hybridomas that were raised against human Lp[a]. Of these, 14 strongly cross-reacted with plasminogen on ELISA screening assays while 16 clearly did not and 10 were only marginally cross-reactive. We took advantage of the homology between plasminogen and apo[a] to define the epitopes of 8 strongly cross-reacting monoclonal antibodies. We were able to subdivide these into four general categories based upon site competition assays (using both plasminogen and Lp[a]), and their reactivity with elastolytically derived plasminogen fragments. Group A monoclonal antibodies (F1 1E3, F2 3A3) recognized epitopes within the kringle 5 and protease domains (miniplasminogen) of plasminogen. The group B monoclonal antibody (F6 1A3) reacted solely with plasminogen kringle 4-like domains and appeared to recognize a limited number of sites on Lp[a]. Group C monoclonal antibodies (F6 1B5, F6 1G9) recognized a second, more frequently distributed site within these kringle 4-like domains. The final group, D, monoclonal antibodies (F6 2C3, F6 2G2, F6 3F4) reacted with a cluster of sites found associated with kringle 4-like domains but also reacted with the miniplasminogen domain. Interestingly, only the members of this group were able to interfere with the proteolytic activity of plasmin. Neither periodate treatment of Lp[a] nor incubation of Lp[a] with epsilon-aminocaproic acid affected the binding of any of our monoclonal antibodies.
Lafferty
M A
MA
University of Delaware, School of Life and Health Sciences, Newark 19716.
Salamon
A M
AM
Usher
D C
DC
D000614:Aminocaproates / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001665:Binding Sites; D001667:Binding, Competitive; D001710:Biotin; D015153:Blotting, Western; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D017270:Lipoprotein(a); D008074:Lipoproteins / Q000276:immunology*; D051379:Mice; D010196:Pancreatic Elastase; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D010504:Periodic Acid / Q000494:pharmacology; D010958:Plasminogen / Q000276:immunology* / Q000378:metabolism
D000614:Aminocaproates; D000911:Antibodies, Monoclonal; D000939:Epitopes; D017270:Lipoprotein(a); D008074:Lipoproteins; D010446:Peptide Fragments; D010504:Periodic Acid; D001710:Biotin; D010958:Plasminogen; D010196:Pancreatic Elastase; D005341:Fibrinolysin