Submitted to: Journal of American Leather Chemists Association
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/10/2004
Publication Date: 7/1/2004
Citation: Mozersky, S.M., Wildermuth, R.J., Marmer, W.N. 2004. Immunochemical estimation of the decorin core-protein content of preparations of bovine decorin. Journal of American Leather Chemists Association. 99(9):280-284. Interpretive Summary: The ability of leather scientists to improve the quality and durability of leather depends on their understanding of the structure of the collagen framework of the hide on a molecular level and their ability to modify that structure during the processing of the hide into the leather product. A key component of the collagen framework involved in both its production and its maintenance is the proteoglycan (PG) decorin. As the designation PG implies, the molecule is part protein ('proteo') and part carbohydrate ('glycan' is a particular type of carbohydrate). This laboratory has been concerned with the possibility of producing softer and more flexible leather by modifying the collagen framework through the controlled removal of some of its decorin. In order to accomplish that, it is necessary for the scientists to be able to measure the amounts of decorin protein and glycan in the hide at various stages before and after it is subjected to treatments designed to remove the PG or one of its components. A procedure to measure the glycan content was recently developed in this laboratory and has been published. An immunochemical procedure to measure the protein (decorin core-protein) content of hide samples has now been developed and is the subject of this communication. By correlating these two measurements of a hide with measurements of the softness and flexibility of leather produced from the hide, it will be possible to design treatments that will optimize the desired properties of the leather product.
Technical Abstract: An indirect sandwich ELISA procedure has been developed for estimation of the decorin core-protein content of preparations of bovine decorin. A mouse anti-bovine decorin antibody is used as the capture antibody, and a rabbit anti-bovine decorin antibody as the detecting antibody. Goat anti-rabbit immunoglobulin G (IgG) labeled with horseradish peroxidase is used to react with the detecting antibody. The peroxidase substrate ortho-phenylenediamine (OPD), when exposed to the (bound) enzyme in the presence of peroxide, yields a soluble product with an absorbance maximum (in acidic medium) at 492 nm. The absorbance of the solution at this wavelength thus reflects the amount of decorin core-protein in the decorin preparation that is recognized by both the capture antibody and the detecting antibody.