CN114062674A - 一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒 - Google Patents
一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒 Download PDFInfo
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Abstract
本发明公开了一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒,包括固定蛋白质,所述固体蛋白质通过表面清洗和表面涂覆处理,①将已知的免疫检查点PD‑1、PD‑L1、CTLA‑4蛋白受体固定上,②根据这些生物分子的特性捕获与其特异性结合的待测患者血清、血浆、淋巴、渗出液、细胞溶液和分泌液中PD‑1、PD‑L1、CTLA‑4,使其充分发生抗原抗体反应。③而后经洗涤和纯化后再进行确认和生化分析。本发明只需一次实验即可检测多种抗体的血清情况,具有省时、高效和高通量平行检测的有点,是现代免疫学技术和蛋白质新材料技术的结晶,且检测结果不仅可通过肉眼定性观察,也可配置阅读仪定量检测,结果能够长期保存,具有重复性好、稳定性和灵敏度高以及特异性强的效果。
Description
技术领域
本发明涉及蛋白质芯片肺癌免疫检测技术领域,尤其是涉及一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒。
背景技术
T细胞是肿瘤免疫答应中的主要效应细胞,在生理情况下T细胞的免疫应答受到共抑制分子即免疫检查点的调节,维持对自身组织的耐受,从而避免自身免疫疾病的发生,共刺激因子CD28与抗原提呈细胞表面的CD28/CD86相互作用,使细胞毒T淋巴细胞相关抗原4(CTLA-4)向胞膜转移,以更强的结合力与CD28/CD86结合,抑制T细胞的活化与增值,程序性死亡受体1(PD-1)在T细胞、自然杀伤细胞(NK)、单核细胞及B细胞整多钟细胞中表达,其功能与CTLA-4相似,程序性死亡配体1(PD-L1)主要在肿瘤细胞表面及肿瘤微环境中表达,肿瘤细胞可异常上调共抑制分子及其相关配体的表达,抑制T淋巴细胞活化,从而导致肿瘤免疫逃逸,随着近年来通过单克隆抗体阻断免疫监测点,如分子PD-1、PD-L1、CTLA-4的研究较为广泛,且研究显示上述分子具有显著的抗肿瘤效应,目前从二线治疗到一线治疗再到联合治疗,免疫检查点抑制剂疗法逐步提高了肺癌治疗的效果,正在改变肺癌的治疗策略。
结合上述,1、在非小细胞肺癌(NSCLC)的二线治疗中,免疫检查点抑制剂已取得突破性进展,PD-1抑制剂Nivolumab用于进展期肺鳞癌和非鳞状 NSCLC 的Ⅲ期临床研究结果显示,Nivolumab组患者的总生存期(OS)、客观缓解率(ORR)、无进展生存期(PFS)及1 年生存率均明显提高。目前,美国 FDA已批准Nivolumab用于以铂类为基础化疗方案的进展晚期( 转移性) 肺鳞癌及非鳞状NSCLC的治疗。PD-1抑制剂 Remborolizunab治疗进展期 NSCLC的Ⅰ期临床研究显示,患者ORR中位缓解持续时间(MDR)明显延长,且PD-L1 高表达,即肿瘤细胞 PD-L1 染色阳性比例(TPS)≥50% 的患者疗效更为显著。基于此结果,美国FDA批准Remborolizunab用于二线治疗PD-L1阳性(TPS≥1%)且在含铂化疗期间或之后发生疾病进展的NSCLC患者。PD-L1抑制剂Atezolizumad治疗NSCLC患者的Ⅲ期临床研究显示,Atezolizumad治疗组中位OS比多西他赛化疗组平均延长了4.2个月。美国FDA批准Atezolizumad用于接受含铂化疗治疗期间或治疗后病情进展以及接受靶向疗法治疗失败的转移性NSCLC患者。
2、Nivolumab单药用于晚期的NSCLC患者一线治疗的Ⅰ期临床研究也取得了一定的成果,但近期公布的Ⅲ期临床研究(CheckMate-026)宣布失败Reck等临床研究首次证实了Pembrolizumad单药一线治疗晚期NSCLC的效果优于标准含铂双药化疗,促使了Pembrolizumab获得美国FDA和欧盟要品监管(EMA)批准,用于治疗PD-L1高表达(TPS≥50%)、表皮生长因子受体(EGFR)及ALK突变阴性的转移性NSCLC患者,langer等研究显示与单纯化疗相比,Keytruda联合化疗的肺癌患者ORR和PFS均有显著提高,基于此项研究,美国FDA加速批准了Keytruda联合化疗一线治疗转移性非鳞状NSCLC,CTLA-4代表抑制剂有Ipilimumab(MD*010)及Tremelimumad,在Ⅱ期临床研究中Ipilimumab二线治疗Ⅵ期NSCLC的结果提示该方案可提高患者的PFS及OS但在后续Ⅲ期临床研究中期分析未显示出临床获益,因而终止了研究,在铂类联合化疗后维持治疗的NSCLCⅡ期临床研究中Tremelimumad不能延长患者PFS。
3、最近的研究表明,肿瘤细胞中错配修复基因缺陷(dMMR)及微卫星不稳定性高(MSI-H)会导致肿瘤出现高水平的突变负荷,产生大量可被免疫系统识别的新抗原,增强体内抗肿瘤免疫活性,进而促进 免疫检查点抑制剂的疗效。因此,美国FDA批准Pembrolizumab用于MSI-H或 dMMR的实体瘤患者,成为 FDA批准的首个依据生物标志物而非肿瘤类型选择的抗肿瘤疗法,标志着免疫治疗正在为肿瘤综合治疗带来巨大变革。
结合上述从二线治疗到一线治疗再到联合治疗,免疫检查点抑制剂疗法逐步提高了肺癌的治疗效果,正在改革肺癌的治疗策略,但是目前只有少部分患者能够应用这种免疫疗法,并且盲目使用会明显提高不良反应的发生率,加速患者恶性进程。究其原因是到目前为止,没有一种有效的检测手段,能对单一个体的肿瘤患者进行其血液中PD-1、PD-L1、CTLA-4含量的检测,以此推断,该病人是否对免疫检查点抑制剂药物有效。因此,开发预测肺癌免疫疗效生物标志物,并用高效的方法筛选可获益人群对实现肿瘤的精准免疫治疗尤为重要。
实用新型内容
有鉴于此,本发明的目的是针对现有技术的不足,提供一种三支点水轮发电机连接装置,可以解决上述背景技术中提出的问题
为达到上述目的,本发明采用以下技术方案:
本发明一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒,包括固体蛋白质,所述固体蛋白质通过表面清洗和表面涂覆处理,①将已知的免疫检查点PD-1、PD-L1、CTLA-4蛋白受体固定上,②根据这些生物分子的特性捕获与其特异性结合的待测患者血清、血浆、淋巴、渗出液、细胞溶液和分泌液中PD-1、PD-L1、CTLA-4,使其充分发生抗原抗体反应。③而后经洗涤和纯化后再进行确认和生化分析。
优选的,所述表面清洗是指反应活性基团通过共价键直接连接于基片表面分子之上形成化学活性单分子层,所述表面涂覆是指通过非共价方式在基片材料表面形成薄膜层。
优选的,所述表面清洗包括基片清洗、硅烷化装饰和偶联其他基因如;(如戊二醛)。
优选的,所述基片清洗分为清洗表面污物和生成一层裸露的羚基层,硅烷化装饰包括氨基、环氧基和巯基化。
本发明的有益效果是:结合背景技术与传统的酶联法、化学发光法、金标法相比较,本发明研发的蛋白质芯片技术检测肺癌免疫检查点检测试剂盒,解决了传统方法一次只能识别一种蛋白的弊端,只需一次实验即可检测多种抗体的血清情况,具有省时、高效和高通量平行检测的有点,是现代免疫学技术和蛋白质血技术的结晶,且检测结果不仅可通过肉眼定性观察,也可通过信号检测进行定量,结果能够长期保存,具体的重复性好、稳定性和灵敏度高以及特异性强的效果。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为本发明的蛋白质芯片反应原理示意图;
具体实施方式
下面结合附图和实施例对本发明作进一步描述。
如图1所示,一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒,结合背景技术与传统的酶联法、化学发光法、金标半点法相比较,本发明研发的蛋白质芯片技术检测肺癌免疫检测试剂盒,解决了传统方法一次只能识别一种蛋白的弊端,只需一次实验即可检测多种抗体的血清情况,具有省时、高效和高通量平行检测的有点,是现代免疫学技术和蛋白质血技术的结晶,且检测结果不仅可通过肉眼定性观察,也可配置阅读仪定量检测,结果能够长期保存,具体的重复性好、稳定性和灵敏度高以及特异性强的效果。
本发明一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒,包括固体蛋白质,所述固体蛋白质通过表面清洗和表面涂覆处理,①将已知的免疫检查点PD-1、PD-L1、CTLA-4蛋白受体固定上,②根据这些生物分子的特性捕获与其特异性结合的待测患者血清、血浆、淋巴、渗出液、细胞溶液和分泌液中PD-1、PD-L1、CTLA-4,使其充分发生抗原抗体反应。③而后经洗涤和纯化后再进行确认和生化分析。
最后说明的是,以上实施例仅用以说明本发明的技术方案而非限制,本领域普通技术人员对本发明的技术方案所做的其他修改或者等同替换,只要不脱离本发明技术方案的精神和范围,均应涵盖在本发明的权利要求范围当中。
Claims (4)
1.一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒,包括固体蛋白质,其特征在于:所述固体蛋白质通过表面清洗和表面涂覆处理,①将已知的免疫检查点PD-1、PD-L1、CTLA-4蛋白受体固定上,②根据这些生物分子的特性捕获与其特异性结合的待测患者血清、血浆、淋巴、渗出液、细胞溶液和分泌液中PD-1、PD-L1、CTLA-4,使其充分发生抗原抗体反应,
③而后经洗涤和纯化后再进行确认和生化分析。
2.根据权利要求1所述的一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒,其特征在于:所述表面清洗是指反应活性基团通过共价键直接连接于基片表面分子之上形成化学活性单分子层,所述表面涂覆是指通过非共价方式在基片材料表面形成薄膜层。
3.根据权利要求2所述的一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒,其特征在于:所述表面清洗包括基片清洗、硅烷化装饰和偶联其他基团如;(如戊二醛)。
4.根据权利要求3所述的一种蛋白质芯片技术检测肺癌免疫检查点检测试剂盒,其特征在于:所述基片清洗分为清洗表面污物和生成一层裸露的羟基层,硅烷化装饰包括氨基化、环氧基化和巯基化。
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114622009A (zh) * | 2022-02-28 | 2022-06-14 | 广州天源高新科技有限公司 | 一种用于早期诊断妊娠糖尿病的miRNA分子标志物及其应用 |
Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1479098A (zh) * | 2002-08-27 | 2004-03-03 | 缪金明 | 高通量IgY抗体芯片构建技术 |
CN1619310A (zh) * | 2004-11-03 | 2005-05-25 | 东南大学 | 肿瘤免疫细胞检测芯片的制备和检测方法 |
CN103033619A (zh) * | 2013-01-08 | 2013-04-10 | 河南生生医疗器械有限公司 | 一种综合检测肺癌标志物的蛋白质芯片试剂盒及方法 |
CN104761745A (zh) * | 2015-03-16 | 2015-07-08 | 北京化工大学 | 一种三维生物芯片基片制备方法 |
CN105527435A (zh) * | 2016-01-06 | 2016-04-27 | 胡海 | 蛋白芯片、蛋白质芯片诊断试剂盒制备及使用方法 |
CN106248955A (zh) * | 2016-09-03 | 2016-12-21 | 长春工业大学 | 一种检测人源化pd‑l1抗体的试剂盒 |
CN109507416A (zh) * | 2018-12-27 | 2019-03-22 | 杭州迪相实业有限公司 | 一种外泌体肿瘤标志物pdl1的快速检测试剂盒 |
CN110456042A (zh) * | 2019-08-09 | 2019-11-15 | 臻悦生物科技江苏有限公司 | 血清学生物标志物在制备试剂盒和/或芯片中的应用、试剂盒或芯片及其制备方法 |
CN111141906A (zh) * | 2020-01-06 | 2020-05-12 | 中南大学湘雅医院 | 一种小细胞肺癌患者外周血循环肿瘤细胞及pd-l1的检测试剂盒 |
CN111381041A (zh) * | 2018-12-29 | 2020-07-07 | 上海细胞治疗集团有限公司 | Pd-1抗体检测试剂盒 |
-
2020
- 2020-07-30 CN CN202010749839.0A patent/CN114062674A/zh active Pending
Patent Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1479098A (zh) * | 2002-08-27 | 2004-03-03 | 缪金明 | 高通量IgY抗体芯片构建技术 |
CN1619310A (zh) * | 2004-11-03 | 2005-05-25 | 东南大学 | 肿瘤免疫细胞检测芯片的制备和检测方法 |
WO2006047928A1 (fr) * | 2004-11-03 | 2006-05-11 | Southeast University | Préparation et applications de puces immunodétectrices pour la détection de cellules cancéreuses |
CN103033619A (zh) * | 2013-01-08 | 2013-04-10 | 河南生生医疗器械有限公司 | 一种综合检测肺癌标志物的蛋白质芯片试剂盒及方法 |
CN104761745A (zh) * | 2015-03-16 | 2015-07-08 | 北京化工大学 | 一种三维生物芯片基片制备方法 |
CN105527435A (zh) * | 2016-01-06 | 2016-04-27 | 胡海 | 蛋白芯片、蛋白质芯片诊断试剂盒制备及使用方法 |
CN106248955A (zh) * | 2016-09-03 | 2016-12-21 | 长春工业大学 | 一种检测人源化pd‑l1抗体的试剂盒 |
CN109507416A (zh) * | 2018-12-27 | 2019-03-22 | 杭州迪相实业有限公司 | 一种外泌体肿瘤标志物pdl1的快速检测试剂盒 |
CN111381041A (zh) * | 2018-12-29 | 2020-07-07 | 上海细胞治疗集团有限公司 | Pd-1抗体检测试剂盒 |
CN110456042A (zh) * | 2019-08-09 | 2019-11-15 | 臻悦生物科技江苏有限公司 | 血清学生物标志物在制备试剂盒和/或芯片中的应用、试剂盒或芯片及其制备方法 |
CN111141906A (zh) * | 2020-01-06 | 2020-05-12 | 中南大学湘雅医院 | 一种小细胞肺癌患者外周血循环肿瘤细胞及pd-l1的检测试剂盒 |
Non-Patent Citations (1)
Title |
---|
朱林霄;黎友伦;: "免疫检查点抑制剂在小细胞肺癌治疗中的临床研究现状", 临床肺科杂志, vol. 25, no. 5, pages 788 - 793 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114622009A (zh) * | 2022-02-28 | 2022-06-14 | 广州天源高新科技有限公司 | 一种用于早期诊断妊娠糖尿病的miRNA分子标志物及其应用 |
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