Immunochemical control of the species origin of porcine crude heparin and detection of ovine and caprine materials.
As a consequence of the outbreak of bovine spongiform encephalopathy (BSE), ruminants materials have been generally banned from the production of heparin. Immunochemical methods have been recently developed for the control of the raw materials used by manufacturers of materials such as porcine mucosa and for the detection of bovine crude heparins. To certify the porcine origin of crude porcine heparins and to exclude ovine or caprine materials, new ELISAs were developed. Rabbit antisera were produced against species-specific antigenic contaminants present in crude heparins or in eluted materials (EM) from the chromatographic step of the purification process. When analysed by line immunoelectrophoresis, these antisera revealed five to eleven antigenic contaminants in the EMs, the major one being the most anodic and predominant antigen in crude heparins. Using the best antisera, competitive indirect ELISAs were optimised. They allowed the detection of porcine, ovine and caprine crude heparins down to a dilution of 0.6 to 1.5 parts per 1000, with CVs ranging from 3 to 12%. These ELISAs complete the set of immunological techniques which can be routinely used by heparin manufacturers to secure their supply chain.
Levieux
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A
INRA, Station de Recherches sur la Viande, Immunochimie, Theix, 63122, Saint-Genès-Champanelle, France.
Rivera
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V
Levieux
D
D
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000941:Antigens / Q000032:analysis*; D002417:Cattle; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006041:Goats; D006493:Heparin / Q000032:analysis* / Q000276:immunology; D007106:Immune Sera / Q000096:biosynthesis; D007122:Immunoelectrophoresis; D011786:Quality Control; D012449:Safety; D012756:Sheep; D013045:Species Specificity; D013552:Swine
D000906:Antibodies; D000941:Antigens; D007106:Immune Sera; D006493:Heparin