Simple ELISA method for the evaluation of soluble HLA class I antigens in human serum

S. Comuzio, F. Puppo, R. Ruzzenenti, A. Orlandini, F. Grillo, S. Brenci, L. Lanza, M. Scudeletti, F. Indiveri

Research output: Contribution to journalArticle

Abstract

A simple sandwich ELISA method has been developed for the quantification of soluble HLA class I antigens (s-HLA) in human serum. The assay utilizes the monoclonal antibody Q6/64, directed to a monomorphic determinant of the HLA α-chain, to capture the antigen and the biotinylated NAMB1 monoclonal antibody, directed to β2-microglobulin, as the detection antibody. The extract of the LG-2 lymphoid cell line and pooled sera from 100 healthy subjects are utilized as standards. The arbitrary value of 100 s-HLA Relative Units/mL (RU/mL) is given to the 1:20 dilution of pooled human sera whose optical density value corresponds to the one of the extract of 1 x 106 LG-2 cells (6.25 μg/mL protein concentration). The assay is easy to perform, specific, reproducible (intra-and inter-assay variations ranging from 3.2% to 8.87%), sensitive (detection limit of 6 RU/mL), and needs a small amount of serum (0.1 mL). The mean serum levels of s-HLA found in 100 healthy normal subjects are 41.9 ± 13.4 RU/mL. The potential uses of the method are discussed.

Original languageEnglish
Pages (from-to)278-283
Number of pages6
JournalJournal of Clinical Laboratory Analysis
Volume5
Issue number4
Publication statusPublished - 1991

    Fingerprint

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Comuzio, S., Puppo, F., Ruzzenenti, R., Orlandini, A., Grillo, F., Brenci, S., Lanza, L., Scudeletti, M., & Indiveri, F. (1991). Simple ELISA method for the evaluation of soluble HLA class I antigens in human serum. Journal of Clinical Laboratory Analysis, 5(4), 278-283.