Identification of decorin and other proteins in bovine hide during its processing into leather

The present work attempts to develop a more accurate and reliable assay for decorin concentration in hides as they are processed to leather. The ELISA technique previously developed for decorin analysis is further improved and optimized by treating the samples with chondroitinase-ABC after dialyzing them in the presence of collagenase. Some newer techniques developed and adapted include SDS-PAGE and Western blotting using immuno-colorimetric staining. Probing the blotted protein with a combination of two different antibodies specific for decorin, i.e., PK1 and 6D6, is more specific and sensitive than using either antibody individually. Western Blotting can be used visually and quantitatively to confirm that ELISA-detected species are indeed decorin. The intact proteoglycan as well as the core protein and resulting fragmentations of decorin are determined after treating the samples with collagenase and chondroitinase ABC alone or a combination of these two enzymes. The fate of the different proteins in raw hides to leather is followed by analyzing the separated protein bands in SDS-PAGE gel using MALDI-TOF peptide mapping.