An in-house ELISA method for measuring surfactant protein A.
An in-house enzyme-linked immunoabsorbant assay (ELISA) for SP-A was successfully developed using in-house polyclonal anti SP-A and a commercial polyclonal anti-rabbit immunoglobulin horseradish peroxidase conjugate system. The standard curve, generated by using 50 ng of SP-A to coat the plate and 1:500 dilution of polyclonal anti SP-A as a primary antibody, was linear for concentrations of SP-A ranging from 4 micrograms/l to 4000 micrograms/l and reproducible. Results of recovery study of SP-A from a known sample of tracheal aspirate ranged from 94%-114%. Intra- and inter-assay coefficients of variations were 2.7% and 5.6% respectively for a known sample of tracheal aspirate. Interference study showed that tracheal aspirate did not interfere with the assay. The assay developed was intended to be used for SP-A measurement in tracheal aspirates obtained from neonates with and without respiratory distress syndrome.
Cheong
K B
KB
Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur.
Cheong
S K
SK
Boo
N Y
NY
Jemilah
M
M
Ton
S H
SH
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D006023:Glycoproteins / Q000097:blood* / Q000276:immunology / Q000592:standards; D006801:Humans; D011510:Proteolipids / Q000097:blood* / Q000276:immunology / Q000592:standards; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000097:blood* / Q000276:immunology / Q000592:standards; D015203:Reproducibility of Results
D006023:Glycoproteins; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants