Simple elisa method for the evaluation of soluble hla class i antigens in human serum
✍ Scribed by Sergio Comuzio; Alessandro Orlandini; Franco Grillo; Dr. Francesco Puppo; Rosaria Ruzzenenti; Sabrina Brenci; Lorella Lanza; Marco Scudeletti; Francesco Indiveri
- Publisher
- John Wiley and Sons
- Year
- 1991
- Tongue
- English
- Weight
- 494 KB
- Volume
- 5
- Category
- Article
- ISSN
- 0887-8013
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✦ Synopsis
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 Q6164, directed to a monomorphic determinant of the HLA a-chain, to capture the antigen and the biotinylated NAMB1 monoclonal antibody, directed to p,-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 UnitsimL (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 l o 6 LG-2 cells (6.25 p,g/mL protein concentration). The assay is easy to perform, specific, reproducible (intraand inter-assay variations ranging from 3.2% to 8.87%), sensitive (detection limit of 6 RUlmL), 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 k 13.4 RUlmL. The potential uses of the method are discussed.