Takeyama M, Yasunaga F, Otaka A, Fujii N, Yajima H
Department of Hospital Pharmacy, Medical College of Oita, Japan.
J Immunol Methods. 1990 Jul 3;130(2):217-22. doi: 10.1016/0022-1759(90)90051-v.
A sensitive and specific microenzyme immunoassay (EIA) procedure for porcine brain natriuretic peptide (BNP)-like immunoreactivity has been developed. Enzyme-labeled antigen was prepared by conjugation of synthetic BNP with beta-D-galactosidase using N-(epsilon-maleimidocaproyloxy)succinimide method. Using a second antibody-coated immunoplate, the minimum amount of BNP-like immunoreactivity (BNP-LI) detectable by this assay system was 1.6 fmol/well. When porcine BNP-LI in porcine plasma was assayed by the present method levels between 1 and 8 pmol/l were detected. Gel filtration of porcine plasma extracts on Sephadex G-25 revealed the presence of two immunoreactive peaks; one eluted at a position identical with that of BNP-26 and the other eluted earlier, close the position of BNP-32.
已开发出一种用于检测猪脑钠肽（BNP）样免疫反应性的灵敏且特异的微酶免疫测定（EIA）方法。使用N-（ε-马来酰亚胺基己酰氧基）琥珀酰亚胺法将合成的BNP与β-D-半乳糖苷酶偶联制备酶标记抗原。使用包被有第二抗体的免疫板，该检测系统可检测到的BNP样免疫反应性（BNP-LI）的最小量为1.6 fmol/孔。当用本方法检测猪血浆中的猪BNP-LI时，检测到的水平在1至8 pmol/l之间。在Sephadex G-25上对猪血浆提取物进行凝胶过滤，发现存在两个免疫反应峰；一个在与BNP-26相同的位置洗脱，另一个洗脱较早，接近BNP-32的位置。
