Method and collagen coated slide for assaying collagenase

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase

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435287, 4353171, 435805, 435810, 530356, C12Q 138, C12M 100

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active

049313860

ABSTRACT:
The collagenolytic activity of mammalian biologic fluids is assayed by obtaining a specimen of such fluid; applying the fluid to a collagen film bonded to a surface such as that of a glass slide; incubating the slide; washing the slide; staining the slide with a dye, and determining the extent of dissolution of the collagen film from the unstained areas of the slide. The slide is preferably prepared by blending insoluble collagen with a solution of HCl to form a dispersion; applying the dispersion to the slide surface; drying the slide; and baking the slide to cross-link the collagen and bond it to the slide surface. The dispersion applied to the slide contains collagen particles having sizes of at least 1 micron and preferably at least 20 microns. The assay procedure is particularly useful for the determination of the collagenase activity of gingival crevicular fluid (GCF), in which case the fluid specimen is obtained on an absorbent medium such as a filter paper strip. An aspect of the invention is the provision of a kit suitable for use in dental offices which includes absorbent strips; one or more slides having collagen bonded to the surface thereof; and a container of a dye solution capable of staining collagen. The kit may also include one or more collagenase standards.

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