Identification of human nonpancreatic-type ribonuclease by antibodies obtained against a synthetic peptide.
An antibody that recognizes human nonpancreatic-type ribonuclease was obtained by immunizing a rabbit with a 14-residue synthetic peptide corresponding to the N-terminal sequence of eosinophil-derived neurotoxin which is identical to human liver ribonuclease. This amino acid sequence is unique to this protein. The anti N-peptide antibody was purified by protein A-Sepharose and by using ELISA and SDS-PAGE immunoblot techniques, the antibody reactivity against EDN and partially purified nonpancreatic-type ribonucleases from human plasma and urine was observed. Cross-reactivity with bovine pancreatic ribonuclease A and other proteins was not detected. In addition, the activity of the nonpancreatic-type ribonuclease was not affected by the antibody. The immune response was elicited without the need for a carrier protein showing that the N-terminal sequence of nonpancreatic ribonuclease contains a specific epitope. This antibody can be used for the immunological identification of both the native and denatured forms of this type of enzyme.
Bravo
M I
MI
Department de Bioquímicai Biologia Molecular, Facultat de Ciències, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Cuchillo
C M
CM
Nogués
M V
MV
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D002417:Cattle; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D047208:Eosinophil-Derived Neurotoxin; D006801:Humans; D015151:Immunoblotting; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000276:immunology; D010455:Peptides / Q000276:immunology*; D012260:Ribonucleases / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification
D009498:Neurotoxins; D010455:Peptides; D047208:Eosinophil-Derived Neurotoxin; D012260:Ribonucleases