title: One-step sandwich enzyme immunoassay for human laminin using monoclonal antibodies.
abstract: A one-step sandwich enzyme immunoassay (one step sandwich EIA) for human serum immunoreactive laminin was set up with a pair of monoclonal antibodies prepared against human placental laminin P1 fragment. The assay was characterized by carrying out two immunoreactions simultaneously, laminin P1 fragment reacting with both a monoclonal antibody as a solid phase and a horseradish peroxidase-labeled monoclonal antibody (Fab') against human laminin P1 fragment as conjugate. Sensitivity of the immunoassay was 0.01 ng/well (0.5 microgram/l), and linearity was obtained between 0.01-20 ng/well (0.5-1,000 micrograms/l). The levels of laminin in sera from normal individuals and patients with liver cirrhosis, hepatocellular carcinoma and primary biliary cirrhosis were 103 +/- 15 micrograms/l, 228 +/- 70 micrograms/l, 341 +/- 163 micrograms/l and 232 +/- 93 micrograms/l, respectively. Protein immunoblotting showed that the serum immunoreactive laminin measured by the assay was a fragment with rel mol mass of 200 kDa.
authors.affiliation: Fuji Chemicals Industries Ltd., Toyama, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Fujimoto
authors.forename: H
authors.initials: H
authors.lastname: Sejima
authors.forename: M
authors.initials: M
authors.lastname: Kyotani
authors.forename: K
authors.initials: K
authors.lastname: Iwata
authors.forename: T
authors.initials: T
authors.lastname: Ichida
authors.forename: A
authors.initials: A
authors.lastname: Ooshima
mesh_terms: D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003094:Collagen / Q000378:metabolism; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D007797:Laminin / Q000097:blood* / Q000276:immunology; D008107:Liver Diseases / Q000097:blood; D010920:Placenta / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D007797:Laminin; D003094:Collagen