title: Simple ELISA method for the evaluation of soluble HLA class I antigens in human serum.
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 alpha-chain, to capture the antigen and the biotinylated NAMB1 monoclonal antibody, directed to beta 2-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 10(6) LG-2 cells (6.25 micrograms/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.
authors.affiliation: S.p.a. Italiana Laboratori Bouty, Milano.
authors.forename: S
authors.initials: S
authors.lastname: Comuzio
authors.forename: F
authors.initials: F
authors.lastname: Puppo
authors.forename: R
authors.initials: R
authors.lastname: Ruzzenenti
authors.forename: A
authors.initials: A
authors.lastname: Orlandini
authors.forename: F
authors.initials: F
authors.lastname: Grillo
authors.forename: S
authors.initials: S
authors.lastname: Brenci
authors.forename: L
authors.initials: L
authors.lastname: Lanza
authors.forename: M
authors.initials: M
authors.lastname: Scudeletti
authors.forename: F
authors.initials: F
authors.lastname: Indiveri
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal; D001769:Blood / Q000276:immunology; D002460:Cell Line; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D015234:HLA-A Antigens / Q000097:blood*; D006801:Humans; D008297:Male; D008875:Middle Aged
chemical_list: D000911:Antibodies, Monoclonal; D015234:HLA-A Antigens