Differentiation of gelatins using polyclonal antibodies raised against tyrosylated bovine and porcine gelatins.
Gelatin is obtained from bones and hides/skin, mainly from cows and pigs using alkaline or acidic processes. The use of bovine gelatin in feed, food, and pharmaceutical products has been restricted by regulatory authorities as a consequence of the outbreak of bovine spongiform encephalopathy (BSE). On the other hand, some religions ban the porcine gelatin for human consumption. Thus, there is a need for methods able to control the species origin of gelatins. The large similarity in structure of gelatins from different origins has made unsuccessful their differentiation by physicochemical methods. Moreover, the development of immunochemical methods has been hampered by the poor immunogenicity of gelatins. We obtained high titers antibodies upon immunization of rabbits with tyrosylated bovine and porcine gelatins. Using indirect and competitive indirect ELISAs we observed large differences in titers and specificity among rabbits and during the course of immunization. Some of the antisera were not sensitive to the species origin of raw material or to the process used for gelatin production and could be used for gelatin quantitation in food. Other antisera detected the porcine acidic gelatins with 10- to 30-fold higher sensitivity than their bovine counterparts and could be used for the differentiation of the species origin of gelatins. Lastly, other antisera were highly sensitive to subtle changes in conformation of gelatins obtained by alkaline or acidic processes such as a 1,000-fold higher reactivity of bovine acid hide gelatin compared to that of its limed counterpart or a 30,000-fold higher reactivity of porcine acid bone compared to that of its limed counterpart; such antisera could be used to monitor the process induced structural changes of collagen during its transformation to gelatin.
Venien
Annie
A
INRA, Theix, SRV-Immunochimie, 63122, Saint-Genès-Champanelle, France.
Levieux
Didier
D
D000143:Acids / Q000737:chemistry; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D001667:Binding, Competitive / Q000276:immunology; D017610:Calcium Compounds / Q000737:chemistry; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005780:Gelatin / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D007114:Immunization; D010087:Oxides / Q000737:chemistry; D011817:Rabbits; D013552:Swine; D014443:Tyrosine / Q000737:chemistry / Q000276:immunology*
D000143:Acids; D000906:Antibodies; D017610:Calcium Compounds; D010087:Oxides; D014443:Tyrosine; D005780:Gelatin; C016538:lime