PO.ET02.05 · 实验与分子治疗
MPSA-AB5000:一种基于细胞的高通量膜蛋白筛选阵列,用于精准的早期脱靶评估以加速更安全抗体治疗药物的开发
MPSA-AB5000: A high-throughput cell-based membrane protein screening array for accurate early-stage off-target assessment to accelerate safer antibody therapeutic development
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
抗体药物开发是一个漫长而复杂的过程,常受脱靶结合的阻碍,脱靶结合影响约25-30%的临床前候选物。随着CD3双特异性抗体、抗体-药物偶联物(ADC)和CAR-T疗法等先进治疗模式日益重要,早期且准确的脱靶评估已变得至关重要。然而,包括免疫组化(IHC)、ELISA和流式细胞术(FACS)在内的传统检测方法,在灵敏度、特异性和通量方面仍存在局限。为解决这些局限,我们开发了膜蛋白筛选阵列(MPSA-AB5000),这是一个基于细胞的高通量平台,用于全面鉴定抗体靶点和脱靶点。MPSA-AB5000在活细胞中表达广泛的人膜蛋白,维持天然构象和翻译后修饰,以确保具有生理相关性的结合分析。MPSA-AB5000是一个基于细胞的高通量平台,用于鉴定抗体靶点和脱靶点,增强抗体特异性筛选并降低开发风险。为提供最高水平的灵敏度,MPSA-AB5000采用荧光素酶报告基因细胞系检测。无洗涤策略减少了假阴性,信号放大则实现了卓越的区分度。此外,该平台整合了基于蛋白A的报告系统,利用蛋白A对Fc的强结合亲和力——尤其是对人IgG1、IgG2和IgG4——将抗体固定在报告细胞上。这使得即使在IgG1-LALA或IgG4等Fc修饰形式中也能检测到脱靶诱导的信号。总体而言,MPSA-AB5000为抗体靶点解卷积和特异性评估提供了一种稳健、多功能且具生理相关性的解决方案,加速了更安全抗体治疗药物的发现与优化。
查看英文原文 English abstract
Antibody drug development is a lengthy and complex process, often hindered by off-target binding that affects approximately 25-30% of preclinical candidates. With the growing importance of advanced modalities such as CD3 bispecific antibodies, antibody-drug conjugates (ADCs), and CAR-T therapies, early and accurate off-target assessment has become essential. However, conventional assays-including immunohistochemistry (IHC), ELISA, and flow cytometry (FACS)-remain limited in sensitivity, specificity, and throughput.To address these limitations, we developed the Membrane Protein Screening Array (MPSA-AB5000), a high-throughput, cell-based platform for comprehensive identification of antibody targets and off-targets. The MPSA-AB5000 expresses a broad spectrum of human membrane proteins in live cells, maintaining native conformation and post-translational modifications to ensure physiologically relevant binding analysis. MPSA-AB5000 is a high-throughput, cell-based platform for identifying antibody targets and off-targets, enhancing antibody specificity screening and reducing development risks. To provide the highest level of sensitivity, MPSA-AB5000 uses luciferase reporter cell line detection. The wash-free strategy reduces false negatives, and signal amplification makes remarkable discrimination. Furthermore, the platform integrates a Protein A-based reporter system, leveraging Protein A's strong Fc-binding affinity-particularly for human IgG1, IgG2, and IgG4-to immobilize antibodies on reporter cells. This allows detection of off-target-induced signaling even in Fc-modified formats such as IgG1-LALA or IgG4. Collectively, the MPSA-AB5000 provides a robust, versatile, and physiologically relevant solution for antibody target deconvolution and specificity evaluation, accelerating the discovery and optimization of safer antibody therapeutics.
利益披露 Disclosure
L. Wang,
Kyinno Biotechnology Co., LTD Employment.
G. Wang,
Kyinno Biotechnology Co., LTD Employment.
Y. Peng,
Kyinno Biotechnology Co., LTD Employment.
J. Ning,
Kyinno Biotechnology Co., LTD Employment.
F. Hao,
Kyinno Biotechnology Co., LTD Employment.