Open AccessDevelopment and validation of an automated particle‐enhanced nephelometric immunoassay method for the measurement of human plasma c1q
Author(s) -
Borque Luis,
Olivan V.,
Iguaz F.
Publication year - 1995
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1860090505
Subject(s) - radial immunodiffusion , immunoassay , chromatography , latex fixation test , chemistry , nephelometry , standard curve , analytical chemistry (journal) , antibody , immunology , medicine
We have developed a sensitive immunoassay based on latex particle agglutination for measuring C1q concentrations in human plasma. In this simple and fast particle‐enhanced immunoassay, we used carboxylated latex particles (diameter 210 nm) covalently coated with F(ab) 2 fragments of anti‐C1q antibodies. These particles are incubated with diluted sample (400‐fold) for 6 min at room temperature, with the resulting agglutination quantified by measuring the change of light‐scatter produced. The assay has been automated on the Behring nephelometer analyzer with a sampling rate of 150 samples/hr. This assay generates a standard curve in the range of 24–775 mg/L, showing intraassay and interassay precision of <8% and <10%, respectively. Dilution linearity was validated throughout the dynamic range of the assay. There were no interferences from bilirubin, Intralipid, haemoglobin, and rheumatoid factor. Results obtained in 45 clinical samples correlated well with those obtained by a commercial radial immunodiffusion method (r=0.936), and with those obtained by the Behring immunoprecipitation nephelometric test (r=0.950). The mean concentration in plasma from healthy subjects was 180 mg/L and the reference interval was from 128 to 237 mg/L. This latex nephelometric procedure is a convenient method and an interesting alternative to other immunoassays for routine measurement of human C1q.