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抗番茄红素单克隆抗体的制备与应用

Generation and Application of Monoclonal Antibody Against Lycopene.

作者信息

Tsibezov Valeriy V, Bashmakov Yuriy K, Pristenskiy Dmitry V, Zigangirova Naylia A, Kostina Ludmila V, Chalyk Natalya E, Kozlov Alexey Y, Morgunova Elena Y, Chernyshova Marina P, Lozbiakova Marina V, Kyle Nigel H, Petyaev Ivan M

机构信息

1 Gamaleya Research Center of Epidemiology and Microbiology , Ministry of Health, Moscow, Russia .

2 Lycotec Ltd , Cambridge, United Kingdom .

出版信息

Monoclon Antib Immunodiagn Immunother. 2017 Apr;36(2):62-67. doi: 10.1089/mab.2016.0046. Epub 2017 Apr 12.

Abstract

A monoclonal antibody (Mab) against lycopene was developed from hybridoma clones obtained from BALB/c mice immunized with trans-isomer of lycopene (t-lycopene, t-LC) conjugated with colloidal gold particles. An alternating immunization schedule which included injection of both formulations of immunogen (without and with Freund's adjuvant) was most effective in the elucidation of a measurable immune response to the t-Lycopene conjugate. Selected hybridoma clones were able to produce an Mab positive in competition assay. In particular, preincubation of 6B9 Mabs with t-LC abolished the ability of 6B9 Mabs to bind LC in the competition assay. Mabs produced by other clones (4F10, 4A3, and 3B12) worked similarly. Analysis of antigen specificity showed that 6B9 Mab raised against t-LC did not recognize other carotenoids such as lutein and carotene. Mab 6B9 was shown to recognize lycopene on a glass surface and in the settings of indirect immunofluorescence experiments performed in cultured hepatocytes and alveolar macrophages incubated with and without lycopene, as well as in sebum and corneocyte specimens from the skin of volunteers supplemented with nutraceutical formulation of lycopene. Newly generated Mabs against lycopene may provide a valuable tool for different analytical assays of lycopene content in various biological, agricultural, and food products.

摘要

一种抗番茄红素的单克隆抗体(Mab)是由从用与胶体金颗粒偶联的番茄红素反式异构体(t-番茄红素,t-LC)免疫的BALB/c小鼠获得的杂交瘤克隆产生的。一种交替免疫方案,包括注射两种免疫原制剂(有无弗氏佐剂),在阐明对t-番茄红素偶联物的可测量免疫反应方面最为有效。选定的杂交瘤克隆能够在竞争试验中产生阳性的单克隆抗体。特别是,6B9单克隆抗体与t-LC预孵育后,在竞争试验中消除了6B9单克隆抗体结合LC的能力。其他克隆(4F10、4A3和3B12)产生的单克隆抗体也有类似作用。抗原特异性分析表明,针对t-LC产生的6B9单克隆抗体不识别其他类胡萝卜素,如叶黄素和胡萝卜素。在间接免疫荧光实验中,6B9单克隆抗体在玻璃表面以及在添加和未添加番茄红素的培养肝细胞和肺泡巨噬细胞中,以及在补充了番茄红素营养制剂的志愿者皮肤的皮脂和角质形成细胞标本中,均显示能识别番茄红素。新产生的抗番茄红素单克隆抗体可能为各种生物、农业和食品产品中番茄红素含量的不同分析检测提供有价值的工具。

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