Spectrophotometric determination of carp vitellogenin using a sequential injection analysis technique equipped with a jet ring cell.
Spectrophotometric determination of carp vitellogenin using a sequential injection analysis technique equipped with a jet ring cell.
复制标题
使用配备喷射环池的顺序注射分析技术对鲤鱼卵黄蛋白原进行分光光度测定。
DOI：
10.1016/s0039-9140(02)00437-x
复制
发表时间：
2002
期刊：
影响因子：
6.1
通讯作者：
S. Okutani
中科院分区：
文献类型：
--
作者：
N. Soh;Hideshi Nishiyama;K. Mishima;T. Imato;T. Masadome;Y. Asano;Y. Kurokawa;H. Tabei;S. Okutani
将抗体固定化微球转移到喷射环（JR）池中，采用顺序注射分析（SIA）技术测定鲤鱼卵黄原蛋白（Vg）。该测定是基于夹心免疫测定法，其中使用抗鲤鱼Vg抗体和鲤鱼Vg之间的两种类型的反应。即，将用于第一反应步骤的抗体固定在微珠（Sephadex珠）上，并且在反应的第二步骤中使用用辣根过氧化物酶（HRP）标记的抗体。过氧化氢和邻苯二胺（OPD）的混合溶液被用作反应中的发色团源。将微球固定抗体、Vg分析物、HRP标记抗体和显色液按程序顺序自动引入SIA系统的JR池中，并利用氧化OPD产物的吸光度来测定存在的Vg量。考虑到对Vg测定的灵敏度，确定第一步和第二步免疫反应的最佳孵育时间分别为120和60 min。在此条件下，氧化OPD的吸光度与Vg浓度之间具有良好的线性关系。在该系统中，测定Vg的检测下限约为5 ng ml− 1。该方法简便、准确，适用于Vg的测定。
A sequential injection analysis (SIA) technique, in which antibody-immobilized microbeads were transferred to a jet ring (JR) cell, was used in determination of carp vitellogenin (Vg). The determination is based on a sandwich immunoassay in which two types of reactions between anti carp Vg antibodies and carp Vg are used. Namely, the antibody for the first reaction step was immobilized on microbeads (Sephadex beads), and an antibody labeled with a horseradish peroxidase (HRP) was used in the second step of the reaction. A mixed solution of hydrogen peroxide and o-phenylenediamine (OPD) was used as the source of the chromophore in the reaction. The microbeads-immobilized antibody, Vg analyte, HRP-labeled anitbody and the color developing solution were introduced automatically into the JR cell of the SIA system in a programmed sequence, and the absorbance of the oxidized OPD product was used to determine the amount of Vg present. The optimal incubation times for the immuno-raction for the first and the second steps were determined at 120 and 60 min, respectively, taking into account the sensitivity to the Vg determination. Under these conditions, a good linear correlation was obtained between Vg concentration and the absorbance of the oxidized OPD. The lower detection limit for the determination of Vg was about 5 ng ml−1in this system. The method developed here represents a simple, accurate method for the determination method of Vg.
停流连续注射分析原理及其在蛋白水解酶痕量动力学测定中的应用。
影响因子：
7.4
作者：
Ruzicka,J;Gübeli,T
通讯作者：
Gübeli,T
射流环池：一种在可再生固体支持物上进行流动注射光谱和显微镜检查的工具。
影响因子：
7.4
作者：
Ruzicka,J;Pollema,CH;Scudder,KM
通讯作者：
Scudder,KM