Abstract
Development of an enzyme-linked immunosorbent assay, using a monoclonal antibody against a2-macroglobulin, for the diagnosis of systemic lupus erythematosus
Cazzolla,Nicolas; Saso,Luciano; Grima,Josephine; Leone,Maria Grazia; Grippa,Eleanora; Cheng,Chuen-yan; Silvestrini,Bruno
Clinical Biochemistry 32(4): 249-255
Publication date: 1999
Objectives
To develop an enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody (mab) directed against abnormally glycosylated serum a-macroglobulin (a-M) from patients with systemic lupus erythematosus (SLE).
Design and methods
Serum a-M purified by HPLC from patients with SLE was injected in a Balb/c, CB6 F1 female mouse and hybrid cell lines were screened using a-M Glu-C fragments derived from SLE and normal donors (NHS). A mab was selected and used to develop an ELISA by which sera from NHS (n = 14), SLE (n = 34), rheumatoid arthritis (n = 15), Sjögren's syndrome (n = 11), mixed connective tissue diseases (n = 12), and liver diseases (n = 11) were analyzed.
Results
The affinity of the mab for a-M from SLE, but not from the other diseases, was higher compared to NHS, as demonstrated by immunoblotting and ELISA.
Conclusions
The ELISA was capable of recognizing changes of glycosylation of a-M in SLE and may be useful for its differential diagnosis.
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