Production and characterization of monoclonal antibodies specific for bovine oviductal glycoproteins

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Abstract
Four monoclonal antibodies (MAbs) were produced in rat cells against materials obtained from the bovine oviduct. The MAbs (1H10, 2A4, 2C5, and 1B12) reacted strongly with the oviductal epithelium of cows in the follicular phase. An immunohistochemical study demonstrated that each of the MAbs bound strongly to the supranuclear cytoplasm of epithelial cells of the oviduct, but not of the stromal cells. Antigens that reacted with the MAbs were characterized by immunoblotting analysis of proteins after fractionation by SDS-PAGE under reducing conditions of oviductal flushings obtained from cows at estrus. All MAbs strongly stained a band of protein with a molecular weight (MW) of 85–97 kD. MAb 1H10 also reacted with the minor band of a 55-kD protein. Similar antigens were not detected in uterine flushings, follicular fluid, and serum. No specific immunohistochemical reactivity with the MAbs was observed with other tissues from the reproductive tract and with nonreproductive tissues. The immunohistochemical reactions were completely eliminated by pretreatment of tissues with trypsin, but not with periodic acid, results that suggest that the antigenic determinants that react with the MAbs were proteinaceous rather than carbohydrate. We have thus established clones that produce 4 MAbs that are specific for oviduct-specific glycoproteins of the cow.