Bravo M I, Cuchillo C M, Nogués M V
Department de Bioquímicai Biologia Molecular, Facultat de Ciències, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Biol Chem Hoppe Seyler. 1995 Sep;376(9):555-60. doi: 10.1515/bchm3.1995.376.9.555.
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.
通过用与嗜酸性粒细胞衍生神经毒素的N端序列相对应的14个残基合成肽免疫兔子，获得了一种识别人类非胰腺型核糖核酸酶的抗体，该序列与人类肝脏核糖核酸酶相同。此氨基酸序列是该蛋白质所特有的。抗N肽抗体通过蛋白A-琼脂糖纯化，并使用ELISA和SDS-PAGE免疫印迹技术观察了该抗体对EDN以及从人血浆和尿液中部分纯化的非胰腺型核糖核酸酶的反应性。未检测到与牛胰腺核糖核酸酶A和其他蛋白质的交叉反应性。此外，该抗体不影响非胰腺型核糖核酸酶的活性。无需载体蛋白即可引发免疫反应，这表明非胰腺核糖核酸酶的N端序列包含一个特定表位。该抗体可用于此类酶天然形式和变性形式的免疫学鉴定。
