Immunoglobulins that bind to uncoated ELISA plate surfaces: Appearance in mice during infection with lactate-dehydrogenase-elevating virus and in human anti-nuclear antibody-positive sera
✍ Scribed by Dr. William A. Cafruny; Daniel P. Heruth; Mary Jo Jaqua; Peter G. W. Plagemann
- Book ID
- 102908329
- Publisher
- John Wiley and Sons
- Year
- 1986
- Tongue
- English
- Weight
- 693 KB
- Volume
- 19
- Category
- Article
- ISSN
- 0146-6615
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✦ Synopsis
Immunoglobulins present in the blood plasma of mice infected with lactatedehydrogenase-elevating virus (LDV) were found to bind strongly in the presence of 0.05 % Tween 20 to the uncoated surfaces of wells of certain ELISA plates with previously recognized high protein-binding capacity. The binding was readily distinguishable from non-specific background binding of immunoglobulins present in normal mouse plasma. The binding components absorbed to protein A and had molecular weights in the 150-300 kDa range. Binding of the purified IgG fraction was progressively inhibited by increasing the concentration of Tween 20 in the diluent and by preincubation of the fraction at pH 3-4 for 10 min. The appearance of plate-binding IgM and IgG during LDV infection corresponded approximately with previously reported time courses of appearance of IgM-and IgG-containing circulating immune complexes and of specific IgM and IgG anti-LDV antibodies in LDV-infected mice. We conclude that complexes of IgG and IgM with LDV antigens have a much higher affinity for ELISA plates with high protein-binding capacity than uncomplexed immunoglobulins. Immune complexes did not significantly bind to ELISA plates with low protein-binding capacity, which, therefore, are suitable for measuring specific antiviral antibodies. Preliminary experiments with human anti-nuclear antibody-positive serum samples demonstrated markedly elevated non-specific binding of immunoglobulins to high-binding-capacity ELISA plates.