CN111679074A - 一种前列腺癌的外周血tcr标志物及其检测试剂盒和应用 - Google Patents
一种前列腺癌的外周血tcr标志物及其检测试剂盒和应用 Download PDFInfo
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Abstract
本发明公开了一种前列腺癌的外周血TCR标志物及其检测试剂盒和应用。该标志物包括序列为SEQ ID NO.1～100所示的蛋白中的至少一种。本发明基于高通量测序方法，只需要采取少量外周血，提取RNA，通过对样本的处理建立免疫图谱文库，再经过高通量测序和TCR数据分析，首先确定前列腺癌外周血中特征性TCR序列，然后将待测样本测试结果与该特征性TCR序列比对，从而确定是否患有前列腺癌。本发明能够同时比较巨大数量的前列腺癌特异性TCR序列，相比单独检测一种或几种标记物，具有更高的特异性和准确性，提高了诊断效率。
Description
技术领域
本发明属于基因工程技术领域，具体涉及一种前列腺癌的外周血TCR标志物及其检测试剂盒和应用。
背景技术
前列腺癌是指发生在前列腺的上皮性恶性肿瘤。根据2004年WHO《泌尿系统及男性生殖器官肿瘤病 理学和遗传学》，前列腺癌病理类型上包括腺癌(腺泡腺癌)、导管腺癌、尿路上皮癌、鳞状细胞癌、腺鳞 癌，其中前列腺腺癌占95％以上，因此，通常我们所说的前列腺癌就是指前列腺腺癌。
前列腺癌发生的主要危险因素包括遗传以及家庭因素等，与生活习惯也有关系。有报道称前列腺癌的 发病与性活动、饮食习惯有关。性活动较多者患前列腺癌的风险增加。高脂肪饮食与发病也有一定关系。 此外，前列腺癌的发病与种族、地区可能有关。前列腺癌发病年龄在55岁前处于较低水平，55岁后逐渐 升高，发病率随着年龄的增长而增长，高峰年龄是70～80岁。家族遗传型前列腺癌患者发病年龄稍早， 年龄≤55岁的患者占43％。
从全球范围来看，发达国家的前列腺癌发病率高于发展中国家，目前美英法德日占据总发患者数的 70％。随着发展中国家的经济高速增长，发展中国家近年来发病率也在快速提升。2012年我国肿瘤登记地 区前列腺癌发病率为9.92/10万，列男性恶性肿瘤发病率的第6位。我国2016年前列腺癌新发患者12万 人，预计2030年，我国前列腺癌新发患者数量将达到23.7万人，新发患者数量的年复合增长率为5％。
早期前列腺癌可以进行有效诊疗和控制，未转移前列腺癌生存时间较长，转移后不可治愈。由于早期 筛查渗透率较低，因此发展中国家前列腺癌死亡率高于发达国家。在美国，患病5年的患者生存率在98％ 以上，而同样的患者在中国生存率仅为50％。以此，寻求更简便更可靠的早期筛查诊断手段，对前列腺癌 的防治具有重大现实意义。
前列腺癌早期常无症状，随着肿瘤的发展，前列腺癌引起的症状可概括为两大类：
1、压迫症状：逐渐增大的前列腺腺体压迫尿道可引起进行性排尿困难，表现为尿线细、射程短、尿 流缓慢、尿流中断、尿后滴沥、排尿不尽、排尿费力，此外，还有尿频、尿急、夜尿增多、甚至尿失禁。 肿瘤压迫直肠可引起大便困难或肠梗阻，也可压迫输精管引起射精缺乏，压迫神经引起会阴部疼痛，并可 向坐骨神经放射。
2、转移症状：前列腺癌可侵及膀胱、精囊、血管神经束，引起血尿、血精、阳痿。盆腔淋巴结转移 可引起双下肢水肿。前列腺癌常易发生骨转移，引起骨痛或病理性骨折、截瘫。前列腺癌也可侵及骨髓引 起贫血或全血象减少。
目前临床诊断前列腺癌的主要方法有：
1、直肠指检：在前列腺癌的早期诊断中极为重要,其准确率可达50％—70％。很多学者主张在前列腺 癌高发地区,对中年以上男性定期进行直肠指检,将使很多病人得到早期诊断及根治的机会。
2、穿刺活检：经会阴、直肠穿刺,取活体组织检查,其诊断的正确率可达70％—80％。
3、影像学检验：经直肠前列腺超声和盆腔MRI检查。
4、前列腺液检验：其阳性率可达90％以上。绝大多数病例可由此得到确诊。
5、血清PSA检验：
前列腺特异抗原(PSA，Prostate Specific Antigen)是前列腺相关的一种抗原。PSA是分子量为32kD 的单链糖蛋白，是一种酷似糜蛋白酶的丝氨酸蛋白酶。他由前列腺上皮细胞分泌产生，正常情况下分泌进 入精液，在精囊包的分裂和精液的液化上发挥生理作用。正常时仅有极低水平的PSA存在于血液中，血清 中PSA浓度的增加预示前列腺发生病理变化或受到创伤。在绝大多数前列腺癌患者中PSA水平升高。
上诉检验方法都具有一定局限性。一方面，因前列腺癌早期无明显症状，或与前列腺增生、前列腺炎 等疾病症状类似，因此很多患者会因抗拒直肠指检等原因，错失早期发现癌变的时机。另一方面，根据美 国的研究发现，利用PSA等方法筛查前列腺癌存在过度诊断和过度治疗的问题。为了改善此状况，2010 年美国国家综合癌症网络制定的《前列腺癌临床实践指南》中首次将严密观察而不是采取“积极治疗”作为 经前列腺穿刺活检确诊为前列腺癌患者的选项之一。因此，亟需一种能更便捷灵敏，且可以追踪前列腺癌 发展状况的筛查、检验方法。
发明内容
针对现有技术中的上述不足，本发明提供一种前列腺癌的外周血TCR标志物及其检测试剂盒和应用， 能准确快速的判断待测样本中是否有较高前列腺癌风险患者。
为实现上述目的，本发明解决其技术问题所采用的技术方案是：
一种前列腺癌的外周血TCR标志物，该标志物包括序列为SEQ ID NO.1～100所示的蛋白中的至少一 种，具体序列见表1。
表1标志物序列
进一步地，标志物的蛋白序列为SEQ ID NO.1～100所示的序列经取代、缺失和/或替换一个或多个碱 基后，能表达相同功能的蛋白。
进一步地，标志物为外周血TCR CDR3序列。
上述标志物在制备治疗前列腺癌的制剂中的应用。
进一步地，制剂中包括含有该标志物的T细胞受体，或能表达产生该标志物的T细胞受体的质粒、病 毒载体或核酸片段。
一种用于前列腺癌检测的试剂盒，包括能与上述标志物发生特异性结合的抗体。
一种制剂，包括能与上述标志物发生特异性结合的抗体；所述制剂可用于对前列腺癌进行诊断、预测、 检测或筛查。
一种检测前列腺癌的蛋白质芯片，该蛋白质芯片包括基片和点样在基片上特异性抗体，该特异性抗体 为能与上述标志物发生特异性结合的抗体。
本发明的原理为：人体内的B淋巴细胞和T淋巴细胞是获得性免疫系统中重要的两类细胞。B细胞通 过细胞表面的B细胞受体(BCR)识别抗原，后期BCR在B细胞分化成浆细胞时，表达成抗体，分泌到 细胞外。T细胞通过细胞表面的T细胞受体(TCR)识别抗原。BCR和TCR的多样性是建立获得性免疫 系统的基础。BCR多样性的理论值是1018，TCR多样性的理论值是1014。BCR与TCR序列中，抗原决定 簇3(CDR3)是决定其抗原特异性最重要的部分，因此CDR3的序列被认为可以代表BCR、TCR序列的 特性。
在各种疾病中，随着不同的抗原刺激，BCR和TCR的多样性或者表达水平都会发生改变。因此，利 用BCR或者TCR高通量测序结果可以追踪疾病的发生、发展。人体内细胞中，衰老蛋白质降解后，其片 段会被运输到细胞表面，通过组织相容性抗原II(MCHII)呈递给免疫系统中的T细胞。正常细胞呈递的 抗原片段，由于免疫耐受的关系，不会引起免疫反应。一旦当正常细胞出现癌变后，突变的基因表达的异 常蛋白，其片段被呈递到细胞表面后，就会引起人体免疫系统产生针对性的免疫反应。因此，分析BCR 或TCR的变化，能够检测出肿瘤的发生和发展。
本发明的有益效果为：
1、本发明中，首先利用1300个非前列腺癌的对照组样本、和10个前列腺癌病人的TCR高通量测序 数据建立了人工智能分析模型，通过和这些前列腺癌特异性TCR序列对比，可以清楚的判断待测样本中 是否有较高前列腺癌风险者。
2、通过高通量测序分析TCR变化可以发现很早期的前列腺癌，利用前列腺癌特有的TCR CDR3序列 分析人的免疫系统中的T细胞对前列腺癌的反应，是一种新型的检测方法。
3、本发明通过采用高通量测序技术同时比较数量巨大的特异性TCR序列，比起单独检测一种或几种 标记物，具有更高的特异性和准确性。
4、本发明中使用的高通量测序仪器成本低于大型影像学设备，且可向第三方外包，此外，采样、处 理的人力成本低于同时检测多种标记物，也低于大量细胞学检测，因此，本发明大大降低了检测成本。
5、本发明只需要采取少量外周血，采样简便、安全。
6、本发明中所述TCR CDR3序列，可用于前列腺癌的免疫治疗。
附图说明
图1为本发明中，对照组CDR3序列及前列腺癌特征序列。横坐标代表某一特定氨基酸组合的CDR3 序列被加入对照序列集合或前列腺癌特征序列集合的先后顺序，纵坐标代表该序列在某一样本中重复出现 次数CX的对数值；前列腺癌患者的免疫图谱具有多个种类且重复次数较高的前列腺癌特征序列，健康人 极少前列腺癌特征序列，而未知受试者的前列腺癌特征比较明显，说明罹患前列腺癌风险较高。
图2为本发明中，针对前列腺癌特征序列集，计算健康人、非肿瘤病人、非前列腺癌肿瘤患者和前列 腺癌患者的前列腺癌特征性指数，健康人、非肿瘤病人、非前列腺癌肿瘤患者的前列腺癌特征性指数均与 前列腺癌患者具有显著差异，证明了前列腺癌特征序列集的特异性。据此可以判断未知受试者是否罹患前 列腺癌。
具体实施方式
下面对本发明的具体实施方式进行描述，以便于本技术领域的技术人员理解本发明，但应该清楚， 本发明不限于具体实施方式的范围，对本技术领域的普通技术人员来讲，只要各种变化在所附的权利要求 限定和确定的本发明的精神和范围内，这些变化是显而易见的，一切利用本发明构思的发明创造均在保护 之列。
实施例1通过免疫图谱分析，获得前列腺癌TCR标志物CDR3序列集
1、采样及免疫图谱分析(方法参考专利ZL201910300069.9)
采集1301名对照组(包括健康人和非肿瘤疾病病人，1300人用于建立模型，1个健康人用于验证)、 11名前列腺癌患者(10人用于建立模型，1人用于验证)及1名未知健康状况受试者的外周血(每人10mL)， 通过高通量测序得到受试者和对照组的TCR的抗原决定簇3(CDR3)氨基酸序列，保证每个样本的功能 性TCR的CDR3序列总数综合不低于30000；
2、对每个样本的TCR的CDR3序列进行随机不放回抽样，使每个样本的CDR3序列数量总和均为 30000。对任一特定CDR3序列X，在单样本测序结果中重复出现次数计为CX；
3、通过分析TCR CDR3数据，确定前列腺癌TCR标志物CDR3序列：
a)将1300名用于建立模型的对照组样本的所有CDR3序列归总去重，设为对照序列集；
b)将10名用于建立模型的前列腺癌样本的所有CDR3序列归总去重，再去除所有与对照序列集中包 含序列重复的序列，设为前列腺癌特征序列集。作图如附图1A所示，其中横坐标代表某一特定氨基酸组 合的CDR3序列被加入对照序列集合或前列腺癌特征序列集合的先后顺序，纵坐标代表该序列在某一样本 中重复出现次数CX的对数值。
c)按照同样作图方法，将1名健康人、1名前列腺癌患者和1名健康状况未知受试者的免疫图谱，参 照对照序列集合和前列腺癌特征序列集合进行作图，见附图1B-D。从图中可见，前列腺癌患者的免疫图 谱中，含有较多种类且较高重复出现次数的前列腺癌特征序列；健康人的免疫图谱中，只有极少量前列腺 癌特征序列；而未知健康状况受试者，有高于健康人的前列腺癌特征序列，说明此人有较高风险罹患前列 腺癌。
d)将前列腺癌特征序列集中，将所有出现在两个及以上参与建模前列腺癌样本里的CDR3序列，按 “所有参与建模前列腺癌样本里该序列单样本中重复出现次数CX的总和×包含该序列的参与建模前列腺 癌样本数”从高到低排序，排名前100者即为前列腺癌TCR标志物CDR3序列，具体序列如SEQ ID NO.1～100所示。
实施例2验证前列腺癌TCR标志物CDR3序列集的特异性
1、采样及免疫图谱分析(方法参考专利ZL201910300069.9)
采集339名非前列腺癌的肿瘤患者、2名未知健康状况受试者的外周血(每人10mL)，通过高通量测 序得到受试者和对照组的TCR的抗原决定簇3(CDR3)氨基酸序列，保证每个样本的功能性TCR的CDR3 序列总数综合不低于30000；对每个样本的TCR的CDR3序列进行随机不放回抽样，使每个样本的CDR3 序列数量总和均为30000。
2、从实施例1的对照组中随机挑选100名健康人及45名非肿瘤疾病病人。
3、根据来自实施例1的100名健康人、45名非肿瘤疾病病人、10名前列腺癌患者，以及实施例2新 获取的339名非前列腺癌肿瘤患者、2名未知健康状况受试者的免疫图谱，分析其前列腺癌特征性指数。
其中前列腺癌特征性指数定义为：某个样本中，属于前列腺癌特征序列集的所有CDR3序列在该样本 内重复出现次数CX的总和。
分析结果见下表2及附图2。前列腺癌组与健康人(p＝1.2E-63)、非肿瘤疾病(p＝6.7E-31)、其它肿瘤 (p＝1.2E-194)都有显著差异，这证明了前列腺癌特征序列集的特异性。
表2不同样本组的前列腺癌特征性指数
4、分析各组的前列腺癌特征指数(表3)，未知健康状况受试者(检测样本)中，2人的前列腺癌特 征指数均高于“其它肿瘤”组的平均值+2×SD(23+2×161＝346)，此2人具有较高风险罹患前列腺癌。与临 床体检结果对照后，2人确为前列腺癌患者。此实施例证明了利用前列腺癌特征序列集及前列腺癌特征性 指数，预测受试者罹患前列腺癌风险的可行性。
表3前列腺癌特征性指数分析
| 健康人 | 非肿瘤疾病 | 其他肿瘤 | 前列腺癌 | 检测样本 | |
| Mean | 8.04 | 7.98 | 23.45 | 8806.30 | 4141.00 |
| SD | 12.45 | 6.84 | 161.38 | 2455.35 | 1159.66 |
| mean+2SD | 32.94 | 21.65 | 346.20 |
综上所述，本发明所述前列腺癌TCR标志物CDR3序列，确实具有显著的前列腺癌特异性，不仅可 以用于前列腺癌预测受试者罹患前列腺癌风险，未来还可用于前列腺癌的生物免疫治疗。
序列表
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<210> 45
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 45
Ala Asn Pro Arg Asp Ser Gln Tyr Gly Tyr Thr
1 5 10
<210> 46
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 46
Ala Ser Ser Leu Thr Leu Arg Ile Phe Glu Thr Gln Tyr
1 5 10
<210> 47
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 47
Ser Val Asp Gly Gly Val Thr Glu Ala Leu
1 5 10
<210> 48
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 48
Ala Ser Ser Glu Gly Ala Thr Asp Asn Glu Gln Phe
1 5 10
<210> 49
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 49
Gln Gln Leu Ser Arg Leu Gly Gln Gly Pro Glu Ile Leu Trp Leu His
1 5 10 15
<210> 50
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 50
Ala Ser Thr Ser Glu Val Gln Glu Thr Gln Tyr
1 5 10
<210> 51
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 51
Ser Ala Pro Asp Glu Arg Leu Tyr Gly Tyr Thr
1 5 10
<210> 52
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 52
Ala Ser Ser Leu Lys Ala Ser Ile His Thr Asp Thr Gln Tyr
1 5 10
<210> 53
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 53
Gln Gln Phe Phe Pro Gly Asp His Arg Tyr Ala Val
1 5 10
<210> 54
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 54
Ala Ser Ser Trp Leu Thr Tyr Glu Gln Phe
1 5 10
<210> 55
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 55
Ala Ser Ser Leu Val Arg Gly Pro Ser Leu Arg Glu Gln Tyr
1 5 10
<210> 56
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 56
Ala Ser Ser Tyr Ser Gly Thr Gly Thr Ala Gly Glu Lys Leu Phe
1 5 10 15
<210> 57
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 57
Ala Ser Ser Glu Lys Arg Glu Ile Asn Glu Gln Phe
1 5 10
<210> 58
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 58
Ala Ser Ile Asn Asp Arg Gly Ala Asn Thr Glu Ala Phe
1 5 10
<210> 59
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 59
Ala Ser Ser Pro Val Pro Ala Glu Gln Tyr
1 5 10
<210> 60
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 60
Ala Ser Ser Val Glu Ser Arg Gln Asn Thr Glu Ala Phe
1 5 10
<210> 61
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 61
Ala Ser Ala Asp Ser Asp Asn Ser Pro Leu His
1 5 10
<210> 62
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 62
Ser Ala Asp Gly Gly Val Thr Glu Ala Phe
1 5 10
<210> 63
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 63
Ala Ser Ser Leu Gly Ser Gly Ser Tyr Met Asp Glu Gln Leu
1 5 10
<210> 64
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 64
Ala Ile Ser Glu Val Gly Gln Gly Ser Tyr Asn Glu Gln Phe
1 5 10
<210> 65
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 65
Ala Ser Ser Asp Ala Gly Thr Ala Glu Leu Phe
1 5 10
<210> 66
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 66
Ala Ser Ser Tyr Lys Gln Asn Glu Gln Tyr
1 5 10
<210> 67
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 67
Ser Ala Arg Val Pro Gly Gln Gly Asn Thr Gly Glu Leu Phe
1 5 10
<210> 68
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 68
Ala Thr Arg Thr Gly Gly Gly His Glu Gln Tyr
1 5 10
<210> 69
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 69
Ala Ser Ser Leu Gly Asn Val Ala Ser Gly Thr Asp Thr Gln Tyr
1 5 10 15
<210> 70
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 70
Ala Thr Ser Glu Arg Met Gly Asp Thr Gln Tyr
1 5 10
<210> 71
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 71
Ala Ser Ser Arg Gly Ser Phe Asn Ser Asn Gln Pro Gln His
1 5 10
<210> 72
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 72
Ala Ser Ser Trp Asn Met Gly Asp Thr Gln Tyr
1 5 10
<210> 73
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 73
Ala Ser Ser Pro Ser Leu Gly Thr Val Arg Ser Glu Gln Phe
1 5 10
<210> 74
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 74
Ser Ala Arg Ile Leu Asp Gly Tyr Thr
1 5
<210> 75
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 75
Ala Ser Ser Arg Ser Gly Leu Gln Tyr
1 5
<210> 76
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 76
Ala Ser Ser Leu Gly Ala Thr Asn Leu Tyr Glu Gln Tyr
1 5 10
<210> 77
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 77
Ala Ser Ser Leu Gly Ser Gly Ser Tyr Met Asp Glu Gln Ser
1 5 10
<210> 78
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 78
Ala Ser Gly Val Glu Gly Thr Ser Gly Asn Glu Gln Phe
1 5 10
<210> 79
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 79
Ser Ala Arg Ser Ser Gly Tyr Pro His Gln Pro Gln His
1 5 10
<210> 80
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 80
Ala Ser Ser Leu Gly Ser Gly Ser Tyr Met Asp Gly Gln Phe
1 5 10
<210> 81
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 81
Ala Ser Ser Leu Asp Leu Thr Ala Ala Gly Asn Thr Ile Tyr
1 5 10
<210> 82
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 82
Ala Ser Gly Gly Ala Arg Gly His Thr Gly Glu Leu Phe
1 5 10
<210> 83
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 83
Ala Ser Thr Thr Arg Glu Gly Ala Lys Leu Phe
1 5 10
<210> 84
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 84
Ala Ser Ser Leu Gly Gly Gly Ser Tyr Met Asp Glu Gln Phe
1 5 10
<210> 85
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 85
Ala Ser Ser Pro Gly Gly Leu Leu Arg
1 5
<210> 86
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 86
Ala Ser Ser Pro Glu Gly Val Thr Asn Glu Gln Phe
1 5 10
<210> 87
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 87
Ala Ser Ser Pro Arg Arg Gly Ser Gly Gly Asn Glu Gln Phe
1 5 10
<210> 88
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 88
Ala Ser Ser Leu Leu Pro Gly Gly Met Ser Glu Gln Tyr
1 5 10
<210> 89
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 89
Ala Ser Ser Gln Glu Gly Arg Asp Gly Thr Asp Thr Gln Tyr
1 5 10
<210> 90
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 90
Ala Ser Ile Leu Thr Ser Gly Arg Val Asp Glu Gln Phe
1 5 10
<210> 91
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 91
Ala Ser Thr Leu Gly Asn Thr Glu Ala Ser
1 5 10
<210> 92
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 92
Ala Ser Ser Ile Asp Phe Asn Glu Lys Leu Phe
1 5 10
<210> 93
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 93
Ala Ser Ser Ser Arg Pro Arg Gly Gly Arg Ser Tyr Asn Glu Gln Phe
1 5 10 15
<210> 94
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 94
Ser Val Ser Arg Thr Gly Ser Glu Gln Tyr
1 5 10
<210> 95
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 95
Ala Ser Ser Met Thr Gly Pro Gly Glu Gln Phe
1 5 10
<210> 96
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 96
Ala Ser Ser Ala Arg Pro Gly Ala Glu Lys Leu Phe
1 5 10
<210> 97
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 97
Ala Ser Ser Glu Gly Thr Asp Ser Ser Asn Gln Pro Gln His
1 5 10
<210> 98
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 98
Ala Ser Ser Ala Gly Arg Ala Ser Gly Tyr Thr
1 5 10
<210> 99
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 99
Ala Ser Ser Leu Gly Ser Gly Gly Tyr Met Asp Glu Gln Phe
1 5 10
<210> 100
<211> 6
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 100
Gln Gln Leu Pro Arg Ala
1 5
Claims (7)
1.一种前列腺癌的外周血TCR标志物，其特征在于，所述标志物包括序列为SEQ IDNO.1～100所示的蛋白中的至少一种。
2.根据权利要求1所述的前列腺癌的外周血TCR标志物，其特征在于，所述标志物的蛋白序列为SEQ ID NO.1～100所示的序列经取代、缺失和/或替换一个或多个碱基后，能表达相同功能的蛋白。
3.权利要求1所述的标志物在制备治疗前列腺癌的制剂中的应用。
4.根据权利要求3所述的应用，其特征在于，所述制剂包括含有该标志物的T细胞受体，或能表达产生该标志物的T细胞受体的质粒、病毒载体或核酸片段。
5.一种用于前列腺癌检测的试剂盒，其特征在于，包括能与权利要求1所述标志物发生特异性结合的抗体。
6.一种制剂，其特征在于，包括能与权利要求1所述标志物发生特异性结合的抗体。
7.一种检测前列腺癌的蛋白质芯片，其特征在于，所述蛋白质芯片包括基片和点样在基片上特异性抗体，所述特异性抗体为能与权利要求1所述标志物发生特异性结合的抗体。
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| PCT/CN2021/101949 WO2022012290A1 (zh) | 2020-07-11 | 2021-06-24 | 一种前列腺癌的外周血tcr标志物及其检测试剂盒和应用 |
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| CN113109564A (zh) * | 2021-03-15 | 2021-07-13 | 成都益安博生物技术有限公司 | 一种急性髓细胞性白血病的外周血tcr标志物及其检测试剂盒和应用 |
| CN113567682A (zh) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | 一种阿尔茨海默病的外周血tcr标志物及其检测试剂盒和应用 |
| WO2022012290A1 (zh) * | 2020-07-11 | 2022-01-20 | 成都益安博生物技术有限公司 | 一种前列腺癌的外周血tcr标志物及其检测试剂盒和应用 |
| WO2022194039A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种急性b淋巴细胞白血病的外周血tcr标志物及其检测试剂盒和应用 |
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| WO2022012290A1 (zh) * | 2020-07-11 | 2022-01-20 | 成都益安博生物技术有限公司 | 一种前列腺癌的外周血tcr标志物及其检测试剂盒和应用 |
| CN113109564A (zh) * | 2021-03-15 | 2021-07-13 | 成都益安博生物技术有限公司 | 一种急性髓细胞性白血病的外周血tcr标志物及其检测试剂盒和应用 |
| WO2022194039A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种急性b淋巴细胞白血病的外周血tcr标志物及其检测试剂盒和应用 |
| CN113567682A (zh) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | 一种阿尔茨海默病的外周血tcr标志物及其检测试剂盒和应用 |
| WO2023000688A1 (zh) * | 2021-07-23 | 2023-01-26 | 成都益安博生物技术有限公司 | 一种阿尔茨海默病的外周血tcr标志物及其检测试剂盒和应用 |
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