PO.ET06.06 · 实验与分子治疗

TM-PAP,随处可用:跨物种、多宿主面板实现快速读出

TM-PAP, everywhere you need it: Cross-species, multi-host panel for fast readouts

海报缩略图:TM-PAP,随处可用:跨物种、多宿主面板实现快速读出
编号 7229 展板 21 时间 4/22 09:00–12:00 区域 Section 18 主讲 Feng Hao, MD;PhD
分会场 Tumor Diagnostics, Prognostics, and Therapeutic Outcomes
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作者与单位 Authors & Affiliations

Yue Huang, Xiaomeng Gou, Yao Peng, Jinying Ning, Feng Hao

Kyinno Biotechnology Co., LTD, Beijing, China

摘要 Abstract

中文摘要
前列腺酸性磷酸酶 (PAP,又称 ACP3) 是一种便捷、可成药的抗原,当以 TM-PAP (膜定位 PAP) 形式展示在细胞表面时,可用于快速的检测方法开发。与临时抗原系统相比,标准化的跨物种 TM-PAP 面板可在不同宿主和物种屏障之间对抗体和小分子形式进行可靠的头对头测试。此类模型可加速机制确认 (结合、内化)、效应生物学 (ADCC/ADCP/CDC)、有效载荷递送 (ADC、RIC) 和分析检测 (qFACS 滴定、IHC 对照),同时通过同基因 ACP3-KO 背景提供干净的阴性对照。我们组建了 15 个工程化模型,涵盖四种物种变体 (人、小鼠、大鼠、食蟹猴),横跨八个宿主谱系:人类癌症和生产者细胞系 (LNCaP、VCaP [混合池]、PC3、HT-1080、293T) 以及生物生产/啮齿类肿瘤学骨架 (CHO-K1、MC38、CT26)。该面板包括 LNCaP-ACP3-KO (同基因阴性对照);PC3、HT-1080、293T、CHO-K1、MC38 和 CT26 中的人 TM-PAP;293T/CHO-K1 中的小鼠、大鼠和食蟹猴 TM-PAP 直系同源物面板;以及当无需单细胞克隆时用于快速筛选的 LNCaP-ACP3 和 VCaP-ACP3。该平台现支持体外和体内工作流程:多个宿主 (如 LNCaP、PC3、HT-1080、MC38、CT26) 已建立用于小鼠异种移植/同系研究,其余细胞系与体内部署兼容或作为系统对照,实现从结合/内化到药理学和疗效的端到端评估。整合了物种匹配的 TM-PAP cDNA 以生成稳定细胞系 (克隆或混合池),通过流式细胞术 (FACS) 验证表达并进行常规质控 (无支原体,适用时经 STR 鉴定)。ACP3-KO 细胞系通过 CRISPR/Cas9 创建。典型读出包括高通量结合 EC₅₀、pH 敏感性内化,以及使用人或啮齿类效应细胞的 Fc 介导功能,外加用于 IHC/ELISA 的分析对照。该面板使团队能够 (i) 跨物种标准化以抗原为中心的筛选,(ii) 在动物实验前降低交叉反应性和基质效应的风险,以及 (iii) 将检测建立时间从数周压缩到数天——将"TM-PAP,随处可用"带入早期发现、CMC 相邻分析和转化方法开发。
查看英文原文 English abstract
Prostatic acid phosphatase (PAP, a.k.a. ACP3) is a convenient, druggable antigen for rapid assay development when displayed on the cell surface as TM-PAP (membrane-localized PAP). Compared with ad-hoc antigen systems, a standardized, cross-species TM-PAP panel enables reliable head-to-head testing of antibody and small-molecule formats across hosts and species barriers. Such models accelerate mechanism confirmation (binding, internalization), effector biology (ADCC/ADCP/CDC), payload delivery (ADC, RIC), and analytical assays (qFACS titering, IHC controls), while providing clean negative controls via isogenic ACP3-KO backgrounds.We assembled 15 engineered models spanning four species variants (human, mouse, rat, cynomolgus) across eight host lineages: human cancer and producer lines (LNCaP, VCaP [pools], PC3, HT-1080, 293T) plus bioproduction/rodent oncology backbones (CHO-K1, MC38, CT26). The panel includes LNCaP-ACP3-KO (isogenic negative control); human TM-PAP in PC3, HT-1080, 293T, CHO-K1, MC38, and CT26; ortholog panels in 293T/CHO-K1 for mouse, rat, and cynomolgus TM-PAP; and LNCaP-ACP3 and VCaP-ACP3 for rapid screening when single-cell cloning is unnecessary. The platform now supports both in vitro and in vivo workflows: multiple hosts (e.g., LNCaP, PC3, HT-1080, MC38, CT26) are established for murine xenograft/syngeneic studies, and the remaining lines are compatible with in-vivo deployment or serve as system controls, enabling end-to-end evaluation from binding/internalization to pharmacology and efficacy.Species-matched TM-PAP cDNAs were integrated to generate stable lines (clones or pools), with expression verified by flow cytometry (FACS) and routine QC (mycoplasma-free, STR-authenticated where applicable). The ACP3-KO line was created by CRISPR/Cas9. Typical readouts include high-throughput binding EC₅₀, pH-sensitive internalization, and Fc-mediated functions using human or rodent effectors, plus analytic controls for IHC/ELISA. This panel lets teams (i) standardize antigen-centric screening across species, (ii) de-risk cross-reactivity and matrix effects before animal work, and (iii) compress assay setup time from weeks to days-bringing “TM-PAP, everywhere you need it” to early discovery, CMC-adjacent analytics, and translational method development.
利益披露 Disclosure
Y. Huang, Kyinno Biotechnology Co., LTD Employment. X. Gou, 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.

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