Enzyme-linked immunosorbant assay in the rheumatological laboratory

Charles, Peter; Maini, R N

doi:10.1007/978-94-011-1670-1_5

Cited by 1 publication

(1 citation statement)

References 31 publications

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“…The optimum concentration of purified antigen and sera dilutions was determined as described previously [ 17]. Polystyrene microtitre plates (Nunc, Copenhagen, Denmark) were coated overnight at 4°C with 100 μl PL‐12 recombinant antigen or GST protein (concentration 2.5 μg/ml) in TBS buffer (10 m M Tris–HCl pH 7.5, 150 m M NaCl).…”

Section: Methodsmentioning

confidence: 99%

Detection of anti-PL-12 autoantibodies by ELISA using a recombinant antigen; study of the immunoreactive region

García-Lozano

GonzÁlez-Escribano

Rodríguez

et al. 1998

Clinical and Experimental Immunology

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SUMMARYAutoantibodies to aminoacyl-tRNA synthetases are highly associated with myositis and detection is important in clinical diagnosis; however, current methods of screening limit its clinical utility. In the present study, alanyl-tRNA synthetase (PL-12) recombinant protein was obtained by immunological screening of a HeLa expression library and used in an ELISA with 22 anti-PL-12 sera, 200 autoimmune sera negative for PL-12 and 100 healthy individual sera. Sensitivity of the method was 95% (21/22) and specificity 100%. Mapping of the immunoreactive region was carried out using three anti-PL-12 sera and different recombinant protein-derived peptides. Results show that the same conformational epitope located within amino acids 730-951 of the PL-12 antigen outside the catalytic region was recognized by the three anti-PL-12 sera tested. We conclude that ELISA using recombinant protein is an effective and useful method for routine screening for anti-PL-12 autoantibodies.

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Section: Methodsmentioning

confidence: 99%