Development and validation of an enzyme-linked immunosorbent assay for the quantification of a recombinant humanized anti-IL-4Rα monoclonal antibody CM310 in serum and its application to pharmacokinetic study in Sprague-Dawley Rats

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry Pub Date : 2024-07-25 DOI:10.1016/j.ab.2024.115623

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引用次数: 0

Abstract

CM310 is a recombinant humanized monoclonal antibody targeting Interleukin (IL)-4 receptor alpha (IL-4Rα). IL-4Rα blockade prevents IL-4 and IL-13 from binding to their receptor, thereby inhibiting downstream signaling pathways that drive Type 2 helper T-cell (Th2) inflammation. CM310 holds potential for treating Th2-related inflammatory diseases, such as asthma, atopic dermatitis and chronic sinusitis with nasal polyposis. In this study, a direct enzyme-linked immunosorbent assay (ELISA) was developed to measure the concentrations of CM310 in rat serum. Seven calibration standards (ranging from 25 to 1600 ng/mL) and three quality controls (70, 500 and 1250 ng/mL) were defined. The limit of detection (LOD), lower limit of quantification (LLOQ) and upper limit of quantification (ULOQ) were 13, 25 and 1600 ng/mL, respectively. The method exhibited excellent precision and accuracy and successfully applied to in vitro serum stability and pharmacokinetic (PK) studies. In conclusion, we have developed and validated a highly sensitive and selective method for measuring CM310 in Sprague-Dawley rats. The development and validation ELISA method met the acceptable criteria, which suggested that these can be applied to quantify CM310, as well as in PK studies.

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开发和验证用于血清中重组人源化抗IL-4Rα单克隆抗体CM310定量的酶联免疫吸附测定及其在Sprague-Dawley大鼠药代动力学研究中的应用。

CM310 是一种针对白细胞介素 (IL)-4 受体α（IL-4Rα）的重组人源化单克隆抗体。阻断 IL-4Rα 可阻止 IL-4 和 IL-13 与其受体结合，从而抑制驱动 2 型辅助性 T 细胞（Th2）炎症的下游信号通路。CM310有望治疗与Th2相关的炎症性疾病，如哮喘、特应性皮炎和慢性鼻窦炎伴鼻息肉。本研究开发了一种直接酶联免疫吸附测定法（ELISA）来测定大鼠血清中 CM310 的浓度。确定了七个校准标准（25 至 1600 ng/mL）和三个质量控制（70、500 和 1250 ng/mL）。方法的检出限（LOD）、定量下限（LLOQ）和定量上限（ULOQ）分别为13、25和1600 ng/mL。该方法具有良好的精密度和准确度，并成功地应用于体外血清稳定性和药代动力学（PK）研究。总之，我们建立并验证了一种高灵敏度和高选择性的方法来测定Sprague-Dawley大鼠体内的CM310。所开发和验证的ELISA方法符合可接受的标准，这表明这些方法可用于CM310的定量以及PK研究。

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来源期刊

Analytical biochemistry 生物-分析化学

CiteScore

5.70

自引率

0.00%

发文量

283

审稿时长

44 days

期刊介绍： The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field. The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology. The journal has been particularly active in: -Analytical techniques for biological molecules- Aptamer selection and utilization- Biosensors- Chromatography- Cloning, sequencing and mutagenesis- Electrochemical methods- Electrophoresis- Enzyme characterization methods- Immunological approaches- Mass spectrometry of proteins and nucleic acids- Metabolomics- Nano level techniques- Optical spectroscopy in all its forms. The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.