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用于免疫测定的氨基、氯甲基和缩醛官能化乳胶颗粒:一项比较研究。

Amino, chloromethyl and acetal-functionalized latex particles for immunoassays: a comparative study.

作者信息

Izquierdo M P Sanz, Martín-Molina A, Ramos J, Rus A, Borque L, Forcada J, Galisteo-González F

机构信息

Laboratorio de Análisis Clínicos, Complejo Hospitalario San Millán-San Pedro, Logroño (La Rioja), Spain.

出版信息

J Immunol Methods. 2004 Apr;287(1-2):159-67. doi: 10.1016/j.jim.2004.01.020.

Abstract

Latex particles with different functionalized surface groups (amino, acetal and chloromethyl) for the covalent linking of protein molecules were synthesized and characterized. Immunopurified anti-ferritin antibodies were then covalently coupled with a mean efficiency rate (protein covalently bound to latex particles with respect to the total amount of protein added) of 60%. The reagents developed were applied to the measurement of serum ferritin concentration in a turbidimetric procedure, showing a good measuring range and a lowest detection limit of 3.5 ng/ml in the case of the amino-modified particles. These immunological reagents were compared with a commercial nephelometric method, showing a good linear correlation in all cases but no transferability in the acetal and chloromethyl latex with additional carboxyl groups, probably due to interference with other serum components. The differences among latex found in this study indicate that it would be necessary to optimize the assay conditions for each type of particle, in order to achieve a maximum immunoreactivity.

摘要

合成并表征了具有不同功能化表面基团(氨基、缩醛和氯甲基)用于蛋白质分子共价连接的乳胶颗粒。然后,将免疫纯化的抗铁蛋白抗体以60%的平均效率率(相对于添加的蛋白质总量,共价结合到乳胶颗粒上的蛋白质)进行共价偶联。所开发的试剂应用于比浊法测量血清铁蛋白浓度,对于氨基修饰的颗粒,显示出良好的测量范围和3.5 ng/ml的最低检测限。将这些免疫试剂与商业比浊法进行比较,在所有情况下均显示出良好的线性相关性,但对于带有额外羧基的缩醛和氯甲基乳胶则没有转移性,这可能是由于与其他血清成分的干扰所致。本研究中发现的乳胶之间的差异表明,有必要针对每种类型的颗粒优化测定条件,以实现最大免疫反应性。

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