Determination of CD59 protein in normal human serum by enzyme immunoassay, using octyl-glucoside detergent to release glycosyl-phosphatidylinositol-CD59 from lipid complex

• Published in: Immunology letters 2003-12, Vol.90 (2-3), p.209-213
• Main Authors: Landi, A.P.G, Wilson, A.B, Davies, A, Lachmann, P.J, Ferriani, V.P.L, Seilly, D.J, Assis-Pandochi, A.I
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; CD59 Antigens - blood; CD59 Antigens - chemistry; CD59 Antigens - immunology; CD59 Antigens - metabolism; Child, Preschool; Complement; ELISA; Enzyme-Linked Immunosorbent Assay - methods; Glucosides - chemistry; Glycosylphosphatidylinositols - chemistry; Glycosylphosphatidylinositols - metabolism; GPI-anchored proteins; Humans; Infant; Polyethylene Glycols - chemistry; Serum CD59
• ISSN: 0165-2478; 1879-0542
• DOI: 10.1016/j.imlet.2003.07.001

In this study we have optimised the enzyme immunoassay (ELISA) to quantify CD59 antigen in human serum or plasma. The glycosyl-phosphatidylinositol (GPI)-linked form of CD59 is known to complex with serum high-density lipoprotein. For ELISA optimisation, therefore, we investigated the effect of detergents, added to the sample diluent, on the determined values of CD59. Values obtained in the presence of octyl-glucoside (OG) for 20 adults aged 18–35 years and 17 children 1–5 years old were, respectively, 33–119ng/ml (mean ± S.D.: 66±22ng/ml) and 37–143ng/ml (76±33ng/ml). These results were higher than those measured without OG and were in contrast with published results showing absence, or eight to nine times lower levels, of the protein in serum. A known range for serum concentrations of CD59 in healthy individuals will establish an important reference point for clinical work and for the investigation of diseases involving the complement membrane attack complex (MAC) and its regulation.