Novel hapten design, highly sensitive monoclonal antibody production, and immunoassay development for rapid screening of illegally added chloramphenicol in cosmetics

IF 1.6 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of immunological methods Pub Date : 2023-12-22 DOI:10.1016/j.jim.2023.113604

Zhaoxiang Wang , Mian Wang , Xiaoxiang Fu , Jingqi Qian , Min Wang , Guiyu Tan

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引用次数: 0

Abstract

Hapten design and synthesis have been regarded as the key factor to generate high-quality antibodies. In the present study, a novel hapten of chloramphenicol was synthesized, characterized and compared with two conventional haptens. The new hapten generated mAb 4B5 showed higher sensitivity and titer than the other two haptens-based mAbs. The haptens synthesized with the structure of chloramphenicol base generated more sensitive antibodies than the hapten with chloramphenicol succinate, and the spacer arm linked to the phenyl group hapten elicited the strongest antibody response. After optimization, a direct competitive enzyme-linked immunosorbent assay (dcELISA) and a lateral flow immunoassay (LFIA), both based on the mAb 4B5, were developed. The dcELISA had a half maximum inhibition concentration of 0.23 ng/mL and the LFIA showed a cutoff value of 5–10 ng/mL. The LFIA was applied to detect illegally-added chloramphenicol samples in anti-acne cosmetics, five out of 19 samples were tested chloramphenicol containing within 10 min, which result was confirmed with the dcELISA and HPLC. The LFIA has an adequate sensitivity and can be used as a point of care diagnostic device for rapidly screening chloramphenicol in cosmetics.

查看原文本刊更多论文

用于快速筛查化妆品中非法添加的氯霉素的新型合体设计、高灵敏度单克隆抗体的生产和免疫测定法的开发

合粒的设计和合成一直被认为是产生高质量抗体的关键因素。本研究合成了氯霉素的一种新型合体素，对其进行了表征，并将其与两种传统合体素进行了比较。与其他两种合酶相比，新合酶生成的 mAb 4B5 显示出更高的灵敏度和滴度。以氯霉素碱结构合成的合体比以氯霉素琥珀酸结构合成的合体产生的抗体更敏感，而与苯基合体相连的间隔臂引起的抗体反应最强。经过优化，开发出了基于 mAb 4B5 的直接竞争酶联免疫吸附测定（dcELISA）和侧流免疫测定（LFIA）。dcELISA 的半最大抑制浓度为 0.23 纳克/毫升，而 LFIA 的临界值为 5-10 纳克/毫升。应用 LFIA 检测祛痘化妆品中非法添加的氯霉素样品，19 个样品中有 5 个在 10 分钟内被检测出含有氯霉素，该结果与 dcELISA 和 HPLC 的结果一致。LFIA 具有足够的灵敏度，可用作快速筛查化妆品中氯霉素的定点诊断设备。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of immunological methods 医学-免疫学

CiteScore

4.10

自引率

0.00%

发文量

120

审稿时长

3 months

期刊介绍： The Journal of Immunological Methods is devoted to covering techniques for: (1) Quantitating and detecting antibodies and/or antigens. (2) Purifying immunoglobulins, lymphokines and other molecules of the immune system. (3) Isolating antigens and other substances important in immunological processes. (4) Labelling antigens and antibodies. (5) Localizing antigens and/or antibodies in tissues and cells. (6) Detecting, and fractionating immunocompetent cells. (7) Assaying for cellular immunity. (8) Documenting cell-cell interactions. (9) Initiating immunity and unresponsiveness. (10) Transplanting tissues. (11) Studying items closely related to immunity such as complement, reticuloendothelial system and others. (12) Molecular techniques for studying immune cells and their receptors. (13) Imaging of the immune system. (14) Methods for production or their fragments in eukaryotic and prokaryotic cells. In addition the journal will publish articles on novel methods for analysing the organization, structure and expression of genes for immunologically important molecules such as immunoglobulins, T cell receptors and accessory molecules involved in antigen recognition, processing and presentation. Submitted full length manuscripts should describe new methods of broad applicability to immunology and not simply the application of an established method to a particular substance - although papers describing such applications may be considered for publication as a short Technical Note. Review articles will also be published by the Journal of Immunological Methods. In general these manuscripts are by solicitation however anyone interested in submitting a review can contact the Reviews Editor and provide an outline of the proposed review.