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• W2024171691 abstract "The IL-1β precursor (proIL-1β) represents a significant component of total IL-1β production in certain cell types such as keratinocytes, fibroblasts and alveolar macrophages. It has been presumed that immunodetection systems for the mature 17 kDa IL-1β can be used interchangeably for the 35 kDa intracellular proIL-1β. However, during attempts to purify alveolar macrophage proIL-1β, we found that conventional enzyme-linked immunoassays (ELISAs) (using antibodies directed against the 17 kDa mature IL-1β) underestimated the amounts of 35 kDa proIL-1β by at least ten-fold compared to detection by Western blot techniques. This difference was due to the fact that ELISAs, with an antigen capture format (i.e., that use more than one epitope), can more readily see these distinct epitopes on mature or partially processed IL-1β than on the proIL-1β molecule. This problem does not occur with the Western blot technique, either because only one antibody is needed and hence there is no stearic blockade of a second epitope or because it denatures 35 kDa proIL-1β during the immobilization step, presumably better exposing epitopes as expressed on mature 17 kDa IL-1β. The problem with the ELISA can be partially corrected by proteolytic removal of the aminoterminus of 35 kDa proIL-1β with neutrophil elastase. More accurate determinations of proIL-1β by ELISA can be made by using 35 kDa proIL-1β as the reference standard (when the 35 kDa proIL-1β is free of lower molecular weight IL-1β). These data suggest that there are conformational differences between the carboxyterminus of 35 kDa proIL-1β and mature 17 kDa IL-1β which may affect immunodetection when using antibodies directed against mature 17 kDa IL-1β." @default.
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• W2024171691 date "1992-01-01" @default.
• W2024171691 modified "2023-10-14" @default.
• W2024171691 title "Sandwich ELISA formats designed to detect 17 kDa IL-1β significantly underestimate 35 kDa IL-1β" @default.
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• W2024171691 doi "https://doi.org/10.1016/0022-1759(92)90178-v" @default.
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