Messeri G, Orlandini A, Pazzagli M
Laboratory of Endocrinology, USL, Firenze, Italy.
J Biolumin Chemilumin. 1989 Jul;4(1):154-8. doi: 10.1002/bio.1170040123.
Isoluminol derivatives (ID) were early employed in the preparation of tracers for immunoassay. Their wide use was mainly due to their high quantum efficiency, low molecular weight, well-known chemical structure, low cost and high stability. Moreover the light efficiency of some ID may be modified by specific binding to the antibody, thus allowing the development of homogenous immunoassays requiring no bound/free separation step. Some ID are commercially available under both the amino terminal and carboxyl terminal form for conjugation to respectively carboxy derivative of steroid and amino residues of protein. The linking reaction can be commonly carried on via active esters chemistry and can be easily accomplished within one day. Steroid conjugates can then be rapidly purified by silica gel thin layer chromatography, and protein conjugates by gel filtration on a short disposable column. In the field of steroid studies, isoluminol derivative conjugates were prepared for the immunoassay of almost all compounds of clinical interest. When dealing with protein, both antigens and antibody were labelled in this way, resulting in highly specific activity tracers for competitive and non-competitive immunoassays. Recently the labelling of streptavidin with amino-buty-ethyl-isoluminol allowed the development of very sensitive immunoassay methods which take advantage of the biotin-avidin system.
异鲁米诺衍生物（ID）早期被用于制备免疫测定的示踪剂。它们的广泛应用主要归因于其高量子效率、低分子量、已知的化学结构、低成本和高稳定性。此外，一些ID的光效率可通过与抗体的特异性结合进行调节，从而使得无需进行结合/游离分离步骤的均相免疫测定得以发展。一些ID以氨基末端和羧基末端形式均可商购，分别用于与甾体的羧基衍生物和蛋白质的氨基残基偶联。连接反应通常可通过活性酯化学进行，且可在一天内轻松完成。甾体偶联物随后可通过硅胶薄层层析快速纯化，蛋白质偶联物则可通过在短一次性柱上进行凝胶过滤纯化。在甾体研究领域，制备了异鲁米诺衍生物偶联物用于几乎所有具有临床意义的化合物的免疫测定。在处理蛋白质时，抗原和抗体均以这种方式进行标记，从而得到用于竞争性和非竞争性免疫测定的高比活性示踪剂。最近，用氨基丁基乙基异鲁米诺标记链霉亲和素使得利用生物素-链霉亲和素系统的非常灵敏的免疫测定方法得以发展。
