中国新药杂志

定量检测人血清中可溶性PD-L1蛋白浓度的方法开发及验证

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高恩惠 ¹^,²^,³ , 张瑞霞 ⁴ , 黄应峰 ⁴ , 满素勤 ²^,³ , 程远国 ¹^,²

中国新药杂志 | 重大新药创制专项巡礼 2021,30(15): 1374-1380

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中国新药杂志 |重大新药创制专项巡礼 2021 , 30 (15) : 1374 -1380

定量检测人血清中可溶性PD-L1蛋白浓度的方法开发及验证

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高恩惠¹^,²^,³, 张瑞霞⁴, 黄应峰⁴, 满素勤²^,³, 程远国¹^,²

作者信息

¹上海工程技术大学，上海　201620

²中国医药工业研究总院国家上海新药安全评价中心，上海　201203

³上海益诺思生物技术股份有限公司，上海　201320

⁴宝船生物医药科技（上海）有限公司，上海　201203

通讯作者:

满素勤，女，博士，主要从事药动学及药效动力学研究。联系电话：（021）60211999-6721，E-mail：sqman@ncdser.com。

程远国，男，研究员，博士生导师，主要从事药动学研究。联系电话：（021）60211999，E-mail：ygcheng@ncdser.com。

作者简介:

高恩惠，女，硕士研究生，主要从事药动学研究。E-mail：1376904453@qq.com。

张瑞霞，女，博士，主要从事生物制药及药理学研究。E-mail：ruixia.zhang@dragonboatbio.com。

Method development and validation for the quantitative detection of soluble PD-L1 protein concentration in human serum

En-hui GAO¹^,²^,³, Rui-xia ZHANG⁴, Ying-feng HUANG⁴, Su-qin MAN²^,³, Yuan-guo CHENG¹^,²

Affiliations

¹Shanghai University of Engineering Science, Shanghai 201620, China

²National Shanghai Center for New Drug Safe Evaluation and Research, China State Institute of Pharmaceutical Industry, Shanghai 201203, China

³Shanghai Innostar Biotech Co., Ltd., Shanghai 201320, China

⁴Dragonboat Biopharmaceutical Co., Shanghai 201203, China

出版时间: 2021-08-15

文章导航

摘要

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目的：

建立一种基于电化学发光法的高灵敏度、高特异性、快速的人血清中可溶性PD-L1（sPD-L1）浓度的定量检测方法，并对该方法进行全面验证。

方法：

基于Meso Scale Discovery（MSD）技术平台，以鼠抗体PD-L1抗体（mouse anti-human PD-L1 antibody）作为捕获抗体，并采用生物素标记的羊抗人PD-L1抗体（biotinylated goat anti-human PD-L1 antibody）作为检测试剂，定量检测人血清中sPD-L1浓度。该研究对检测方法的关键参数进行了优化，并对该方法的定量范围、准确度和精密度、回收率、稳定性、稀释线性、选择性等性能进行方法学验证。

结果：

建立了高灵敏度、高特异性的快速检测人血清中sPD-L1的MSD方法并完成方法学验证。该方法定量范围为9.375~1 200 pg·mL^-1，精密度、准确度、回收率、稀释线性及选择性良好，样品室温放置4.3 h，稀释处理后实验台放置4.3 h，冷藏冰箱（2 ℃~8 ℃）存放24.4 h，经冻融3次，超低温冰箱（-90 ℃~-70 ℃）放置101 d稳定。

结论：

经过方法学验证，该方法可以用于血清中sPD-L1浓度的定量分析，为PD-1/PD-L1免疫检查点相关药物的临床研究药效学评价提供理论依据，在个性化治疗和伴随诊断领域也有潜在的应用价值。

关键词

可溶性PD-L1 / 电化学发光技术 / 方法学 / 验证

Abstract

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Objective:

To establish a high sensitivity, high specificity and rapid method for the determination of soluble PD-L1 (sPD-L1) concentration in human serum based on electrochemiluminescence, and to verify the method to meet the comprehensive purpose of a detection method.

Methods:

Based on the Meso Scale Discovery (MSD) technology platform, mouse anti-human PD-L1 antibody was used as the capture antibody, and biotinylated goat anti-human PD-L1 antibody was used as the detection reagent to quantitatively detect the concentration of sPD-L1 in human serum. The key parameters of the detection method was optimezed and the quantitative range, accuracy and precision, recovery, stability, dilution linearity, selectivity and other properties of the method were validated.

Results:

A high-sensitivity, high-specificity MSD method for rapid detection of sPD-L1 in human serum was established and the methodology was verified. The quantitative range of this method was 9.375~1 200 pg·mL^-1, and the precision, accuracy, recovery, dilution linearity and selectivity are satisfactory. The samples remained stable placed at room temperature for 4.3 h, at test bench for 4.3 h after dilution treatment, and at the refrigerator (2 ℃~8 ℃) for 24.4 h, freeze-thaw for 3 times, and in an ultra-low temperature refrigerator (-90 ℃~-70 ℃) for 101 days.

Conclusion:

After validation, it suggested that this method could be used to quantitatively analyze the concentration of sPD-L1 in serum to provide a theoretical basis for the pharmacodynamic evaluation of PD-1/PD-L1 immune checkpoint related drugs in clinical research. It also has potential applications in personalized therapy and concomitant diagnosis.

Key words

soluble PD-L1 / Meso Scale Discovery / methodology / validation

引用本文

高恩惠, 张瑞霞, 黄应峰, 满素勤, 程远国. 定量检测人血清中可溶性PD-L1蛋白浓度的方法开发及验证. 中国新药杂志, 2021 , 30 (15) : 1374 -1380 .

En-hui GAO, Rui-xia ZHANG, Ying-feng HUANG, Su-qin MAN, Yuan-guo CHENG. Method development and validation for the quantitative detection of soluble PD-L1 protein concentration in human serum[J]. Chinese Journal of New Drugs, 2021 , 30 (15) : 1374 -1380 .

基金

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国家“重大新药创制”科技重大专项资助项目(2020ZX09201-013)
上海市研发公共服务平台项目(18DZ2290100)
浦东新区科技发展基金资助(PKX2019-S09)

参考文献

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参考文献引证文献

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2021年第30卷第15期

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首发时间：2026-10-28
出版时间：2021-08-15

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出版历史

录用日期：2020-12-01

基金

国家“重大新药创制”科技重大专项资助项目(2020ZX09201-013)

上海市研发公共服务平台项目(18DZ2290100)

浦东新区科技发展基金资助(PKX2019-S09)

作者信息

¹上海工程技术大学，上海　201620

²中国医药工业研究总院国家上海新药安全评价中心，上海　201203

³上海益诺思生物技术股份有限公司，上海　201320

⁴宝船生物医药科技（上海）有限公司，上海　201203

通讯作者:

满素勤，女，博士，主要从事药动学及药效动力学研究。联系电话：（021）60211999-6721，E-mail：sqman@ncdser.com。

程远国，男，研究员，博士生导师，主要从事药动学研究。联系电话：（021）60211999，E-mail：ygcheng@ncdser.com。

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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