检测条件对酶联免疫吸附测定抗DNA抗体的影响。

Influence of assay conditions on ELISA determinations of anti-DNA antibodies.

作者信息

Pisetsky D S, Semper K F

出版信息

J Immunol Methods. 1984 Nov 30;74(2):217-27. doi: 10.1016/0022-1759(84)90289-8.

PMID:6209337

Abstract

The influence of assay conditions on anti-DNA determinations by an enzyme-linked immunosorbent assay (ELISA) was investigated to evaluate the detection of various DNA antigenic specificities. Among 4 monoclonal anti-DNA antibodies of MRL-lpr/lpr strain origin, 2 showed higher titers in 100 mM NaCl-50 mM Tris, pH 7.5 (Tris-NaCl) than in phosphate-buffered saline (PBS). The determination of 'polyspecificity' for these monoclonal products also depended on the set of conditions used for assay with inhibitory activity of polynucleotides differing in the 2 buffers. The buffer effects were not confined to the monoclonal antibodies as increases in anti-DNA titers were demonstrated for certain SLE patient sera when assayed in Tris-NaCl rather than PBS; sera of MRL-lpr/lpr mice showed an opposite effect, however, with enhancement of anti-DNA activity by PBS. These results suggest that the representation of antigenic sites on DNA may be variably affected by the conditions of assay, altering quantitative and qualitative assessment of this important serological marker.

摘要

研究了检测条件对酶联免疫吸附测定（ELISA）法检测抗DNA的影响，以评估对各种DNA抗原特异性的检测。在源自MRL-lpr/lpr品系的4种单克隆抗DNA抗体中，有2种在100 mM NaCl - 50 mM Tris，pH 7.5（Tris - NaCl）中的效价比在磷酸盐缓冲盐水（PBS）中更高。这些单克隆产品“多特异性”的测定也取决于用于检测的条件组合，不同缓冲液中多核苷酸的抑制活性不同。缓冲液的影响并不局限于单克隆抗体，因为某些SLE患者血清在Tris - NaCl中检测时抗DNA效价升高，而在PBS中检测时则相反；然而，MRL-lpr/lpr小鼠的血清表现出相反的效果，PBS可增强其抗DNA活性。这些结果表明，DNA上抗原位点的表现可能会因检测条件而受到不同程度的影响，从而改变对这一重要血清学标志物的定量和定性评估。