One-step sandwich enzyme immunoassay for human laminin using monoclonal antibodies.
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.
Fujimoto
N
N
Fuji Chemicals Industries Ltd., Toyama, Japan.
Sejima
H
H
Kyotani
M
M
Iwata
K
K
Ichida
T
T
Ooshima
A
A
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
D000911:Antibodies, Monoclonal; D007797:Laminin; D003094:Collagen