蛋白质的过渡金属羰基标记。一种使用傅里叶变换红外光谱的固相双位点免疫测定新方法。

Transition metal carbonyl labeling of proteins. A novel approach to a solid-phase two-site immunoassay using Fourier transform infrared spectroscopy.

作者信息

Varenne A, Salmain M, Brisson C, Jaouen G

机构信息

Ecole Nationale Supérieure de Chimie de Paris, France.

出版信息

Bioconjug Chem. 1992 Nov-Dec;3(6):471-6. doi: 10.1021/bc00018a002.

DOI:10.1021/bc00018a002

PMID:1463776

Abstract

Labeling of bovine serum albumin (BSA) and anti-human thyroid stimulating hormone (hTSH) monoclonal antibodies (mAbs) was performed using (N-succinimidyl 4-pentynoate)hexacarbonyldicobalt (NSCo2(CO)6). Conditions of coupling were different depending on the protein to be labeled, denaturation of the mAbs occuring with high percentages of organic solvent in the reaction mixture. The influence of reaction time and initial concentration of NSCo2(CO)6 was examined. They were both shown to affect the final coupling rate of the metal carbonyl probe. Preservation of the immunoreactivity toward 125I-hTSH was observed for five conjugates having different NSCo2(CO)6: mAb molar ratios when compared to unmodified and peroxidase-labeled mAbs. Finally, a preliminary study of the quantitative detection of the metal carbonyl mAbs on microtiter wells was achieved using Fourier transform infrared spectroscopy.

摘要

使用（N-琥珀酰亚胺基4-戊炔酸酯）六羰基二钴（NSCo₂(CO)₆）对牛血清白蛋白（BSA）和抗人促甲状腺激素（hTSH）单克隆抗体（mAb）进行标记。偶联条件因待标记的蛋白质而异，反应混合物中高比例的有机溶剂会导致单克隆抗体变性。研究了反应时间和NSCo₂(CO)₆初始浓度的影响。结果表明，二者均会影响金属羰基探针的最终偶联率。与未修饰的和过氧化物酶标记的单克隆抗体相比，观察到具有不同NSCo₂(CO)₆:单克隆抗体摩尔比的五种缀合物对¹²⁵I-hTSH的免疫反应性得以保留。最后，利用傅里叶变换红外光谱法对微量滴定板上的金属羰基单克隆抗体进行了定量检测的初步研究。