You are here: Home :: Area NEM :: Medical science :: Human medicine

Boris Vaisman, Paolo Santambrogio, Paolo Arosio, Eitan Fibach, Abraham M. Konijn

An ELISA for the H-Subunit of Human Ferritin which Employs a Combination of Rabbit Poly- and Mice Monoclonal Antibodies and an Enzyme Labeled Anti-Mouse-IgG

We describe a sensitive ELISA for measuring the H-type subunit of human ferritin. A high detection sensitivity was attained by the use of antibodies from different species and an enzyme-conjugated secondary antibody. It consisted of a sandwich assay using a solid phase coated with a rabbit polyclonal antibody for human ferritin from term placenta and a soluble monoclonal antibody for human H-ferritin, followed by a secondary anti-mouse immunoglobulin (Ig)G conjugated to ?-galactosidase. The assay was calibrated with purified recombinant human H-ferritin from E. coli. The colorigenic chlorophenol red ?-D-galactopyranoside and the fluorogenic 4-methyl-umbelliferyl-?-D-galactopyranoside substrates were used with similar outcome. The described method permits the measurement of human H-ferritin at a concentration ranging from 0.1 to 100 ?g/l (or 20–20000 pg per 200 ?l sample) and is accurate at a concentration as low as 0.3 ?g/l. The coefficient of variation of the assay was 6.05–10.3 % and the recovery of H-ferritin added to cell lysates was 105.8 ± 7.52 %. Depending on the H-ferritin content of the cell line tested, only 600 to 60 000 cells of different human cell lines were needed to measure their H-territin content.

Clinical Chemical Laboratory Medicine, Walter de Gruyter

Print ISSN: 1434-6621
Volume: 37, 02/1999
Pages: 121 - 125

Show full article (external site)

Show all available items of this journal

 

Area NEM