Myelin proteolipid protein (PLP) as a marker antigen of central nervous system contaminations for routine food control.
Spreading transmissible spongiform encephalopathies (TSE) have been widely attributed to transmission by ingestion of mammalian central nervous system (CNS) tissue. Reliable exclusion of this epidemiological important route of transmission relies on an effective surveillance of food contamination. Here, myelin proteolipid protein (PLP) is identified as a specific and largely heat-resistant marker for detection of food contaminations by CNS tissue. PLP is a component of oligodendritic glial sheaths of neuronal processes that is specifically expressed in the CNS. A highly selective polyclonal antibody was developed directed against an epitope present in the full-length PLP protein, but absent from the developmentally regulated splice variant DM-20. In combination with a hydrophobic extraction of PLP from tissue samples, the antibody reliably detected PLP from spinal cord, cerebellum, and cortex of different mammalian species. Consistent with earlier reports on PLP expression, no cross-reactivity was observed with peripheral nerve or extraneural tissue, except for a very faint signal obtained with heart. When applied to an artificial CNS contamination present in sausages, the antibody reliably detected a low concentration (1%) of the contaminant. Application of heat, as used during conventional sausage manufacturing, led to a predominant alteration of arginine residues in the PLP protein and a partial loss of immunoreactivity. In contrast, a stretch of hydrophilic amino acids(112-122) proved to be heat-resistant, preserving the immunogenicity of this PLP epitope during heating. Taken together, the excellent CNS specificity of PLP immunodetection and the presence of a heat-resistant epitope have permitted the development of a highly sensitive immunoassay for CNS contamination in routine food control.
Villmann
Carmen
C
Institut für Biochemie, Emil-Fischer-Zentrum, Friedrich-Alexander-Universität Erlangen-Nürnberg, Fahrstrasse 17, 91054 Erlangen, Germany.
Sandmeier
Barbara
B
Seeber
Silke
S
Hannappel
Ewald
E
Pischetsrieder
Monika
M
Becker
Cord-Michael
CM
D000818:Animals; D000918:Antibody Specificity; D000941:Antigens / Q000032:analysis / Q000276:immunology; D015415:Biomarkers / Q000032:analysis; D002490:Central Nervous System / Q000737:chemistry*; D004355:Drug Stability; D005506:Food Contamination / Q000032:analysis*; D006358:Hot Temperature; D007118:Immunoassay; D018991:Myelin Proteolipid Protein / Q000032:analysis* / Q000276:immunology; D013045:Species Specificity
D000941:Antigens; D015415:Biomarkers; D018991:Myelin Proteolipid Protein