Takeyama M, Mori K, Takayama F, Kondo K, Kitagawa K, Fujii N
Department of Hospital Pharmacy, Medical College of Oita, Japan.
Chem Pharm Bull (Tokyo). 1990 Dec;38(12):3494-6. doi: 10.1248/cpb.38.3494.
A sensitive and specific double-antibody enzyme immunoassay (EIA) for a substance P (SP)-like immunoreactive substance (SP-IS) was developed. For competitive reactions, the SP-antibody was incubated with SP standard (or sample) and beta-D-galactosidase labeled Tyr8-SP (delayed addition). Free and antibody-bound enzyme hapten were separated by using an anti-rabbit immunoglobulin G coated immunoplate. Activity of the enzyme on the plate was fluorometrically determined. The present immunoassay allows detection of 0.4 to 10 fmol/well of SP. Using the present EIA, SP-ISs in human saliva and plasma were determined. The level of SP-IS in human saliva was about 7 pmol/l, which was almost three times higher than that in human plasma.
开发了一种用于P物质(SP)样免疫反应性物质(SP-IS)的灵敏且特异的双抗体酶免疫测定法(EIA)。对于竞争性反应,将SP抗体与SP标准品(或样品)以及β-D-半乳糖苷酶标记的Tyr8-SP一起孵育(延迟添加)。使用抗兔免疫球蛋白G包被的免疫板分离游离的和与抗体结合的酶半抗原。通过荧光法测定板上酶的活性。本免疫测定法可检测到0.4至10 fmol/孔的SP。使用本EIA测定了人唾液和血浆中的SP-IS。人唾液中SP-IS的水平约为7 pmol/l,几乎是人血浆中水平的三倍。