将转基因鸡生物反应器中的重组单克隆抗体应用于酶联免疫吸附试验。

IF 1.7 4区 医学 Q3 PHARMACOLOGY & PHARMACY Biological & pharmaceutical bulletin Pub Date : 2024-01-01 DOI:10.1248/bpb.b24-00175
Takehiro Mukae, Kyoko Yoshii, Isao Oishi
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引用次数: 0

摘要

转基因鸡生物反应器可高效生产含有大量重组蛋白质的蛋清。我们之前开发的转基因鸡能产生针对表皮生长因子受体 2(HER2)的重组单克隆抗体(mAbs)。然而,从转基因鸡蛋中提取的 mAbs 的实际应用尚未得到研究。因此,我们旨在评估这些重组 mAbs 能否用于酶联免疫吸附试验(ELISA)。将转基因鸡蛋中的重组 HER2 mAbs 溶解在磷酸盐缓冲盐水中,然后作为固定抗体直接应用于 96 孔微孔板,无需纯化。使用来自转基因鸡蛋的未纯化重组 HER2 mAbs 的 ELISA 效果与使用市售纯化重组 HER2 mAbs 的 ELISA 效果相当。此外,使用转基因鸡蛋中未纯化的重组 HER2 mAbs 进行的 ELISA 分析显示了很高的抗原特异性,并成功地应用于来自 HER2 阳性和 HER2 阴性细胞系的培养细胞裂解物样本。转基因鸡蛋中未纯化的重组 HER2 mAbs 还能有效地用作纸基 ELISA 中的固定抗体。总之,我们的研究结果表明,转基因鸡蛋中的重组 mAbs 有潜力用于开发经济的 ELISA 装置。据我们所知,这项研究是首次将转基因鸡蛋中的重组 HER2 mAbs 用于 ELISA。
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Application of Recombinant Monoclonal Antibodies from Transgenic Chicken Bioreactors in Enzyme-Linked Immunosorbent Assay.

Transgenic chicken bioreactors can efficiently produce egg whites containing large quantities of recombinant proteins. We previously developed transgenic chickens that produce recombinant monoclonal antibodies (mAbs) against epidermal growth factor receptor 2 (HER2). However, the practical applications of mAbs derived from transgenic eggs have not yet been examined. Therefore, we aimed to evaluate whether these recombinant mAbs can be used in enzyme-linked immunosorbent assay (ELISA). Recombinant HER2 mAbs from transgenic eggs were dissolved in phosphate-buffered saline and applied directly to 96-well microplates as immobilized antibodies without purification. The performance of ELISA using the unpurified recombinant HER2 mAbs from transgenic eggs was comparable to that of ELISA using commercially available purified recombinant HER2 mAbs. Moreover, ELISA using unpurified recombinant HER2 mAbs from transgenic eggs demonstrated high antigen specificity and was successfully applied to samples from cultured cell lysates derived from HER2-positive and HER2-negative cell lines. The unpurified recombinant HER2 mAbs from transgenic eggs were also efficiently used as immobilized antibodies in paper-based ELISA. In conclusion, our findings suggest that recombinant mAbs from transgenic eggs have the potential to be used to develop economic ELISA devices. To the best of our knowledge, this study is the first to use recombinant HER2 mAbs from transgenic eggs in ELISA.

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来源期刊
CiteScore
3.50
自引率
5.00%
发文量
247
审稿时长
2 months
期刊介绍: Biological and Pharmaceutical Bulletin (Biol. Pharm. Bull.) began publication in 1978 as the Journal of Pharmacobio-Dynamics. It covers various biological topics in the pharmaceutical and health sciences. A fourth Society journal, the Journal of Health Science, was merged with Biol. Pharm. Bull. in 2012. The main aim of the Society’s journals is to advance the pharmaceutical sciences with research reports, information exchange, and high-quality discussion. The average review time for articles submitted to the journals is around one month for first decision. The complete texts of all of the Society’s journals can be freely accessed through J-STAGE. The Society’s editorial committee hopes that the content of its journals will be useful to your research, and also invites you to submit your own work to the journals.
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