Development of enzyme-linked immunosorbent assay for human cardiac myosin light chain I.
We have developed a new enzyme-linked immunosorbent assay (ELISA) for the determination of cardiac myosin light chain I (MLI) in human serum. The detection range of ELISA was between 1 and 50 ng/ml, and serial dilutions of human serum showed good linearity. The recovery of different concentrations of cardiac MLI ranged from 87.5% to 100.0%. The intra-assay (n=5) and inter-assay (n=5) showed good results (C.V.<10%). The cross-reactivity with skeletal-myosin light chain (ML) was rather high, but was negligible with other myosin light chains. The concentrations of cardiac MLI in human serum determined by ELISA were similar to those determined by immunoradiometric assay (IRMA). The total assay time and sample volume required for ELISA were approx. 2 h and 25 microl, respectively, while those for IRMA are approx. 24 h and 100 microl. Our novel ELISA method therefore has significant advantages for clinical analysis.
Miyazaki
S
S
Immunology Laboratory, Diagnostics Department, Yamasa Corporation, Choshi, Chiba, Japan. miyazaki@yamasa.com
Hamaoki
M
M
Nagata
A
A
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D009203:Myocardial Infarction / Q000378:metabolism; D018994:Myosin Light Chains / Q000032:analysis*; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D018994:Myosin Light Chains; C076659:myosin light chain I; D006735:Horseradish Peroxidase