CN111693701A - 一种大肠癌的外周血tcr标志物序列及其检测试剂盒和应用 - Google Patents
一种大肠癌的外周血tcr标志物序列及其检测试剂盒和应用 Download PDFInfo
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Abstract
本发明公开了一种大肠癌的外周血TCR标志物及其检测试剂盒和应用,该标志物包括序列SpecSeq1‑100所示蛋白中的至少一种。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,首先确定大肠癌外周血中特征性TCR序列,然后将待测样本测试结果与该特征性TCR序列比对,从而确定是否患有大肠癌。本发明能够同时比较巨大数量的大肠癌特异性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,尤其涉及一种大肠癌的外周血TCR标志物及其检测试剂盒和应用。
背景技术
大肠癌,又称结肠直肠癌(Carcinoma of Colon and Rectum)是胃肠道中常见的恶性肿瘤,在世界范围内是第三位常见的恶性肿瘤,年发病率约100万人,年死亡约50万人。大肠癌也是我国最常见的恶性肿瘤之一,其发病率和病死率在消化系统恶性肿瘤中仅次于胃癌、食管癌和原发性肝癌。在我国恶性肿瘤发病率中居第三位(29/10万),死亡率居第五位(14/10万),而且近年来发病率和死亡率均呈上升趋势。在2012年诊断的全球136万例大肠癌患者中,中国大肠癌新发病例数达25.3万例,占18.6%。而2015中国癌症统计数据则显示,我国大肠癌发病率、死亡率在全部恶性肿瘤中均居第5位,其中新发病例37.6万例,死亡病例19.1万例。中国已成为全球大肠癌每年新发病例数最多的国家。
大肠癌多发生在中年以上的男性,以40~70岁最为多见,但20世纪末发现30岁以下者亦不少见。男女两性发病比例约为2:1。大肠癌可以发生在结肠或直肠的任何部位,但以直肠、乙状结肠最为多见,其余依次见于盲肠、升结肠、降结肠及横结肠。癌瘤大多数为腺癌,少数为鳞状上皮癌及粘液癌。
其发病原因仍不清楚,但有报道认为与饮食习惯相关。高脂、高蛋白、低纤维素饮食会增加患大肠癌的风险。在20%-30%的大肠癌患者中,遗传因素起着重要作用,大肠癌患者的家族成员发生大肠癌的风险增高。慢性溃疡性结肠炎、家族性结肠息肉、遗传性腺瘤病患者,发生大肠癌的概率增高。
随着癌肿的增大,大肠癌的临床表现为排便习惯改变、便血、腹泻、腹泻与便秘交替、局部腹痛等症状,晚期则表现贫血、体重减轻等全身症状。但因为早期症状不明显,多数患者发现时已属中晚期。大肠癌治疗的关键在于早期发现、及时诊断和手术根治。一般癌肿只限于肠壁者预后较好,浸润到肠外者预后较差,年轻患者、癌瘤浸润广泛、有转移者或有并发症者预后不良。
当前对大肠癌主要的检测诊断方法有:
1、影像学检查:
1)纤维结肠镜:诊断结肠癌的重要方法,可以通过肉眼进行形态学诊断,并进行活检。
2)钡剂灌肠:诊断大肠癌诊断的传统方法,在病灶定位上具有优势,但易漏诊较小的肿瘤。
3)CT仿真结肠镜:无创伤性,相对简单安全,适用于无法接受结肠镜检查的患者,尤其是恶性肿瘤引起肠腔狭窄的患者,但无法取得病灶组织进行病理检查。
4)腔内B超、CT、MRI:适用于明确肿瘤的局部浸润、淋巴结转移和远处转移,可完善大肠癌的分期。
2、病理学或细胞学检查:
1)直肠指检:早期诊断的重要方法,但无法发现和诊断结肠部位肿瘤。
2)活检:包括纤维结肠镜活检标本和手术患者的术后标本,适用于早期病变,或狭窄部位活检有困难时。
3、生物化学或分子生物学检验:
1)CEA(CarcinoEmbryonic Antigen,癌胚抗原)和CA199(Carbohydrate Antigen199,糖类抗原199):较为广泛应用在大肠癌的辅助诊断中,但由于在大肠癌中的敏感性和特异性不够理想,因此目前不推荐用于进行大肠癌的筛查。
2)其它分子标志物:包括RAS、BRAF、PIK3CA、MSI等,大量研究报道表面,这些标志物与大肠癌的预后相关,可以用作治疗效果的评估。
上述检验方法都具有一定局限性。一方面,因大肠癌早期无明显症状,或与痔疮、胃肠炎等下消化道疾病症状类似,因此很多患者会因抗拒直肠指检、肠镜需要麻醉等原因,错失早期发现癌变的时机。另一方面,根据美国的研究发现,已知肿瘤标志物对大肠癌缺乏敏感性和特异性,不能作为筛查和早期诊断标准。因此,亟需一种能更便捷灵敏,且可以追踪大肠癌发生、发展状况的筛查、检验方法。
发明内容
本发明的目的在于:针对现有技术中存在的不足,提供一种大肠癌的外周血TCR标志物及其检测试剂盒和应用。
本发明采用的技术方案如下:
一种大肠癌的外周血TCR标志物,包括序列SpecSeq1-100所示蛋白中的至少一种,如表1所示:
表1标志物序列
进一步地,标志物的蛋白序列为SpecSeq1-100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。
上述的标志物在制备检测或治疗大肠癌的制剂中的应用。
进一步地,制剂包括含有该标志物的T细胞受体的质粒、病毒载体或核酸片段。
一种用于大肠癌检测的试剂盒,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;制剂可用于对大肠癌进行诊断、预测、检测或筛查。
一种检测大肠癌的蛋白质芯片,蛋白质芯片包括基片和点样在基片上特异性抗体,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。
本发明应用时,采用基于高通量测序分析外周血TCR序列方法以检测是否患有大肠癌,具体步骤如下:
(1)采集受试者和对照组外周血,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,在该样本中重复出现次数计为CX;
(3)TCR CDR3数据分析:通过分析确定大肠癌TCR标志物CDR3序列:
a)将对照组样本所有CDR3序列归总、去重,设为对照序列集;
b)将大肠癌样本所有CDR3序列归总、去重,再去除所有与对照序列集中包含CDR3序列重复的序列,设为大肠癌特征序列集;
c)将大肠癌特征序列集中,出现于两个及以上样本中的CDR3序列,按“在所有大肠癌样本中该序列重复出现次数CX的总和×包含该序列的大肠癌样本数”从高到低排序,以筛选出大肠癌TCR标志物CDR3序列。
(4)利用大肠癌特征性指数判断未知受试者罹患大肠癌风险:
a)根据健康对照组、非肿瘤病人、非大肠癌肿瘤患者、大肠癌患者以及未知健康状况受试者的免疫图谱,分析其大肠癌特征性指数;
其中大肠癌特征性指数定义为:某个样本中,属于大肠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;
b)当未知健康状况受试者的大肠癌特征性指数高于或接近“其它肿瘤”组的平均值+2×SD时,此人具有较高风险罹患大肠癌;如其大肠癌特征指数接近健康人或非肿瘤疾病组平均值,则表明其患大肠癌风险低。
综上所述,由于采用了上述技术方案,本发明的有益效果是:
1、本发明中,首先利用非大肠癌的对照组样本和大肠癌病人的TCR高通量测序数据建立了人工智能分析模型,通过和这些大肠癌特异性TCR序列对比,可以清楚的判断待测样本中是否有较高大肠癌风险者;
2、通过高通量测序分析TCR变化可以发现很早期的大肠癌,利用大肠癌特有的TCRCDR3序列分析人的免疫系统中的T细胞对大肠癌的反应,是一种新型的检测方法;
3、本发明中,由于采用高通量测序技术,同时比较巨大数量的特异性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,也可用于大肠癌的免疫治疗。
附图说明
为了更清楚地说明本发明实施例的技术方案,下面将对实施例中所需要使用的附图作简单地介绍,应当理解,以下附图仅示出了本发明的某些实施例,因此不应被看作是对范围的限定,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相关的附图。
图1为本发明中,对照组CDR3序列及大肠癌特征序列。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大肠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;大肠癌患者的免疫图谱具有多个种类且重复次数较高的大肠癌特征序列,健康人极少大肠癌特征序列,而未知受试者的大肠癌特征比较明显,说明罹患大肠癌风险较高。
图2为本发明中,针对大肠癌特征序列集,计算健康人、非肿瘤病人、非大肠癌肿瘤患者和大肠癌患者的大肠癌特征性指数,健康人、非肿瘤病人、非大肠癌肿瘤患者的大肠癌特征性指数均与大肠癌患者具有显著差异,证明了大肠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大肠癌。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白,以下结合附图及实施例,对本发明进行进一步详细说明。应当理解,此处所描述的具体实施例仅用以解释本发明,并不用于限定本发明,即所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。因此,以下对在附图中提供的本发明的实施例的详细描述并非旨在限制要求保护的本发明的范围,而是仅仅表示本发明的选定实施例。基于本发明的实施例,本领域技术人员在没有做出创造性劳动的前提下所获得的所有其他实施例,都属于本发明保护的范围。
以下结合实施例对本发明的特征和性能作进一步的详细描述。
实施例1
通过免疫图谱分析,获得大肠癌TCR标志物CDR3序列集:
1、采样及免疫图谱分析
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)、101名大肠癌患者(100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,确定大肠癌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100名用于建立模型的大肠癌样本的所有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,设为大肠癌特征序列集。如图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大肠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名大肠癌患者和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大肠癌特征序列集合进行作图,如图1B-D。从图中可见,大肠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大肠癌特征序列;健康人的免疫图谱中,只有极少量大肠癌特征序列;而未知健康状况受试者,有高于健康人的大肠癌特征序列,说明此人有较高风险罹患大肠癌。
d)将大肠癌特征序列集中,将所有出现在两个及以上参与建模大肠癌样本里的CDR3序列,按“所有参与建模大肠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大肠癌样本数”从高到低排序,排名前100者即为大肠癌TCR标志物CDR3序列。
实施例2
验证大肠癌TCR标志物CDR3序列集的特异性:
1、采样及免疫图谱分析
采集358名非大肠癌的肿瘤患者、3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、100名大肠癌患者,以及实施例2新获取的358名非大肠癌肿瘤患者、3名未知健康状况受试者的免疫图谱,分析其大肠癌特征性指数。
其中大肠癌特征性指数定义为:某个样本中,属于大肠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及图2。大肠癌组与健康人(p=6.2E-120)、非肿瘤疾病(p=1.6E-73)、其它肿瘤(p=3.2E-275)都有显著差异,这证明了大肠癌特征序列集的特异性。
表2不同样本组的大肠癌特征性指数
4、分析各组的大肠癌特征指数(下表3),未知健康状况受试者(检测样本)中,3人的大肠癌特征指数都高“其它肿瘤”组的平均值+2×SD(54+2×95=243),此3人具有较高风险罹患大肠癌。与临床体检结果对照后,3人确为大肠癌患者。此实施例证明了利用大肠癌特征序列集及大肠癌特征性指数,预测受试者罹患大肠癌风险的可行性。
表3大肠癌特征性指数分析表
健康人 | 非肿瘤疾病 | 其他肿瘤 | 大肠癌 | 检测样本 | |
Mean | 43.08 | 31.22 | 53.87 | 14390.62 | 4997.00 |
SD | 21.88 | 16.55 | 94.89 | 2673.29 | 1687.69 |
mean+2SD | 86.84 | 64.32 | 243.64 |
综上所述,本发明所述大肠癌TCR标志物CDR3序列,确实具有显著的大肠癌特异性,不仅可以用于大肠癌预测受试者罹患大肠癌风险,未来还可用于大肠癌的生物免疫治疗。
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等,均应包含在本发明的保护范围之内。
序列表
<110> 成都益安博生物技术有限公司
<120> 一种大肠癌的外周血TCR标志物序列及其检测试剂盒和应用
<160> 100
<170> SIPOSequenceListing 1.0
<210> 1
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Ala Gly Thr Gly Ile Ala Ser Ala Gly Glu Leu Phe
1 5 10
<210> 2
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 2
Ser Val Thr Glu Ser Leu Asp Thr Gln Tyr
1 5 10
<210> 3
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Ala Ser Ser Gln Arg Phe Arg Pro Gln His
1 5 10
<210> 4
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 4
Ala Ser Ser Glu Gly Gln Gly Ser Ile Ile Glu Thr Gln Tyr
1 5 10
<210> 5
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 5
Ser Ala Arg Gly Arg Leu Ala Ala Lys Pro Tyr Glu Gln Tyr
1 5 10
<210> 6
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Ala Ser Ser Gly Phe Thr Thr Asn Gln Pro Gln His
1 5 10
<210> 7
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 7
Ala Ser Ser Phe Gly Asp Gln Gly Asp Glu Gln Tyr
1 5 10
<210> 8
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 8
Ala Ile Ser Arg Ala Thr Val Arg Ala Asp Thr Gln Tyr
1 5 10
<210> 9
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 9
Ala Thr Met Gly Thr Glu Leu Leu Ser Asn Ile Gln Tyr
1 5 10
<210> 10
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 10
Ala Ser Ser Pro Leu Ala Ala Gly Glu Gln Tyr
1 5 10
<210> 11
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 11
Ser Val Glu Ile Ser Thr Ser Gly His Pro Leu Leu Ser Asn Glu Gln
1 5 10 15
Phe
<210> 12
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Ala Ser Ser Gln Lys Gly Thr Gly Thr Leu His
1 5 10
<210> 13
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Ala Ser Ser Val Glu Leu Arg Ala Gly Glu Gly Asn Glu Gln Phe
1 5 10 15
<210> 14
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Ala Ser Ser Phe Thr Arg Ser Val Ser Tyr Glu Gln Tyr
1 5 10
<210> 15
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Ala Thr Ser Ser Val Ala Gly Glu Ile Gln Tyr
1 5 10
<210> 16
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
Ala Thr Ser Arg Asp Asp Gly Phe Glu Gln Phe
1 5 10
<210> 17
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Ala Ser Ser Pro Tyr Pro Gly Tyr Ser Thr Asp Thr Gln Tyr
1 5 10
<210> 18
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Ala Ile Ser Glu Ser Ile Ala Ser Gly Glu Glu Thr Gln Tyr
1 5 10
<210> 19
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Ser Ala Ser Ser Ala Leu Ser Phe Tyr Asn Glu Gln Phe
1 5 10
<210> 20
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Ala Ser Ser Val Ile Gln Gly Phe Tyr Gly Tyr Thr
1 5 10
<210> 21
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 21
Ala Ser Ser Ser Pro Gly Gln Gly Pro Ser Asn Tyr Gly Tyr Thr
1 5 10 15
<210> 22
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Ala Ser Ser Leu Ser Gln Pro Ser Gln Tyr Glu Gln Tyr
1 5 10
<210> 23
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 23
Ala Ile Ser Asp Arg Tyr Arg Gly Asn Thr Gly Glu Leu Phe
1 5 10
<210> 24
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
Ser Ala Arg Ser Trp Thr Gly Arg Ile Ile Asp Glu Gln Phe
1 5 10
<210> 25
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 25
Ala Ser Ser Glu Ala Arg Arg Ala Leu Asn Glu Gln Phe
1 5 10
<210> 26
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Ala Ser Ser Pro Ser Gly Val Trp Tyr Gly Tyr Thr
1 5 10
<210> 27
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Ala Ile Ser Arg Asp Glu Thr Glu Ala Phe
1 5 10
<210> 28
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Ala Ser Ser Leu Ala Leu Ser Trp Pro Gln His
1 5 10
<210> 29
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 29
Ala Ser Arg Pro Thr Ser Gly Trp Val Asp Glu Gln Phe
1 5 10
<210> 30
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Ser Val Ala Ile Asp Glu Arg Gly Gly Ile Glu Gln Phe
1 5 10
<210> 31
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
Ala Ser Ser Phe Thr Ser Asn His Glu Gln Tyr
1 5 10
<210> 32
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Ala Ser Arg Gly Glu Glu Met Gly Gly Thr Asp Thr Gln Tyr
1 5 10
<210> 33
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 33
Thr Glu Gln Gly Ala Gly Thr Glu Ala Phe
1 5 10
<210> 34
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Ser Ala Leu Pro Trp Thr Gly Asp Thr Glu Ala Phe
1 5 10
<210> 35
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 35
Ala Ser Ser Phe Thr Ser Arg Met Glu Phe
1 5 10
<210> 36
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Ala Ser Ser Pro Gly Thr Ser Gly Pro Gly Val Thr Asp Thr Gln Tyr
1 5 10 15
<210> 37
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 37
Ala Ile Ser Val Ser Gln Asp Ile Gly Thr Gln Tyr
1 5 10
<210> 38
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 38
Ala Ser Ser Leu Ala Pro Gly Asp Arg Gly Tyr Asn Glu Gln Phe
1 5 10 15
<210> 39
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 39
Ala Ser Ser Gln Phe Gly Glu Glu Thr Val Glu Thr Gln Tyr
1 5 10
<210> 40
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Ala Thr Ser Ser Gly Gly Thr Ser Gly Lys Trp Glu Gln Phe
1 5 10
<210> 41
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 41
Ala Trp Glu Asp Pro Asn Tyr Gly Tyr Thr
1 5 10
<210> 42
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 42
Ala Ser Ser His Ile Gln Thr Ser Asp Arg Asp Gly Tyr Thr
1 5 10
<210> 43
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 43
Ser Ala Ile Pro Ser Glu Gly Ser Asp Glu Gln Phe
1 5 10
<210> 44
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 44
Ala Ser Ser Pro His Gly Arg Leu Asn Thr Glu Ala Phe
1 5 10
<210> 45
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 45
Ala Trp Ser Pro Trp Asp Ser Gly Asn Tyr Gly Tyr Thr
1 5 10
<210> 46
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 46
Ser Ala Arg Asp Ile Tyr Ser Thr Ser Gly Ser Tyr Glu Gln Tyr
1 5 10 15
<210> 47
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 47
Ala Ser Ser Gln Arg Ala Thr Gly Gly Arg Tyr Thr Glu Ala Phe
1 5 10 15
<210> 48
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 48
Ala Ser Ser Tyr Lys Gly Gly Ile Lys Gly Tyr Glu Gln Phe
1 5 10
<210> 49
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 49
Ser Ala Ala Ala Tyr Ser Gly Glu Gly Pro Asn Glu Gln Phe
1 5 10
<210> 50
<211> 6
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 50
Ala Ser Ser Leu Pro Phe
1 5
<210> 51
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 51
Ser Ala Tyr Arg Pro Gly Leu Ala Gly Gly Lys Asn Tyr Asn Glu Gln
1 5 10 15
Phe
<210> 52
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 52
Ala Ser Trp Ala Gly Gly Gln Ala Phe
1 5
<210> 53
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 53
Gln Gln Tyr Gly Gly Ala Gly Arg Tyr Ala Val
1 5 10
<210> 54
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 54
Ser Val Gly Ser Ala Gly Thr Pro Pro Pro
1 5 10
<210> 55
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 55
Ala Ser Ser Pro Ile Gly Asp Ala Asn Asp Glu Gln Phe
1 5 10
<210> 56
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 56
Ala Ile Ser Glu Ser Thr Phe Ala Ser Glu Glu Thr Gln Tyr
1 5 10
<210> 57
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 57
Ala Ser Thr Asn Lys Arg Asn Glu Gln Phe
1 5 10
<210> 58
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 58
Ala Ser Ser Thr Phe Glu Glu Arg Ser Gly Asn Thr Ile Tyr
1 5 10
<210> 59
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 59
Ala Ser Ser Lys Gly Pro Pro Gly Gln Gly Glu Asp Thr Gln Tyr
1 5 10 15
<210> 60
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 60
Ala Ser Ser Gln Gln Ile Thr Gly Asp Leu Leu Tyr Gly Tyr Thr
1 5 10 15
<210> 61
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 61
Ala Ser Glu Glu Thr Gly Tyr Asp Gly Asn Thr Glu Ala Phe
1 5 10
<210> 62
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 62
Ala Ser Ser Ala Gln His Leu Asn 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 Gly Gln Gly Arg Tyr Thr Glu Ala Phe
1 5 10
<210> 64
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 64
Ala Ser Ser Leu Glu Gly Asn Ser Pro Ser Tyr Glu Gln Tyr
1 5 10
<210> 65
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 65
Ala Ser Ser Arg Pro Gln Gly Ala Leu Ile Tyr Glu Gln Phe
1 5 10
<210> 66
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 66
Ala Ser Ser Leu Tyr Lys Lys Gly Thr Glu Ala Phe
1 5 10
<210> 67
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 67
Ala Ser Lys Phe Ala Met Gly Asn Gln Pro Gln His
1 5 10
<210> 68
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 68
Ala Ser Ser Ile Trp Gly Gln Ser Thr Asp Thr Gln Tyr
1 5 10
<210> 69
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 69
Ala Ser Ser Leu Glu Arg Arg Gly Lys Asp Gly Tyr Thr
1 5 10
<210> 70
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 70
Ala Ile Ser Asp Asp Ser Leu Glu Thr Gln Tyr
1 5 10
<210> 71
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 71
Ala Ser Ser Gln Asp Ser Arg Val Ser Gly Arg Asp Glu Gln Tyr
1 5 10 15
<210> 72
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 72
Ser Ala Ser Pro Met Thr Gly Pro Thr Tyr Glu Gln Tyr
1 5 10
<210> 73
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 73
Ala Gly Met Gly Leu Thr Gly Ala Asn Val Leu Thr
1 5 10
<210> 74
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 74
Ala Thr Ser Ser Arg Asp Gly Ser Gln Glu Thr Gln Tyr
1 5 10
<210> 75
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 75
Ala Ile Ser Pro Pro Gly Gln Gly Gly Ser Glu Gln Phe
1 5 10
<210> 76
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 76
Ala Thr Ser Trp Arg Pro Gly Leu Ala Val Asn Asn Glu Gln Phe
1 5 10 15
<210> 77
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 77
Ala Ser Ser Tyr Ser Gly Thr Gly Ala Val Thr Glu Ala Phe
1 5 10
<210> 78
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 78
Ala Ser Ser Gln Asp Gly Ala Glu Gln Glu Thr Gln Tyr
1 5 10
<210> 79
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 79
Ala Ser Ser Glu Asn Lys Ala Tyr Thr
1 5
<210> 80
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 80
Ala Thr Ser Asp Leu Trp Gly Pro Leu Lys Gly Glu Gln Phe
1 5 10
<210> 81
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 81
Ala Ser Ser Pro Pro Asp Met Pro Tyr Glu Gln Phe
1 5 10
<210> 82
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 82
Ala Ser Gly Gln Ala Tyr Gly Asp Thr Glu Ala Phe
1 5 10
<210> 83
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 83
Ser Ala Arg Gly Pro Leu His Arg Gly Arg Thr Asp Thr Gln Tyr
1 5 10 15
<210> 84
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 84
Ala His Arg Ser Arg Gly Asp Glu Gln Tyr
1 5 10
<210> 85
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 85
Ala Ser Ser Ser Gly Gln Asp Glu Phe
1 5
<210> 86
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 86
Ser Val Glu Trp Gly Phe Gly Gln Glu Thr Gln Tyr
1 5 10
<210> 87
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 87
Ser Ala Ser Asp Arg Asp Asn Tyr Gly Tyr Thr
1 5 10
<210> 88
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 88
Ser Ala Arg Gln Leu Asp Gly Thr Ser Ala Tyr Glu Gln Phe
1 5 10
<210> 89
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 89
Ala Ser Thr Ala Asp Gly Gln Gln Tyr Asn Glu Gln Phe
1 5 10
<210> 90
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 90
Ala Ser Ser Glu Ser Thr Ser Gly Arg Glu Glu Glu Thr Gln Tyr
1 5 10 15
<210> 91
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 91
Ala Ile Val Met Gly Asn Thr Ile Tyr
1 5
<210> 92
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 92
Ala Ser Ser Gln Glu Gly Thr Gly Gly Lys Ala Gln His
1 5 10
<210> 93
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 93
Ala Trp Ser Pro Lys Gly Arg Lys Ala Phe
1 5 10
<210> 94
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 94
Ala Ser Ser Gln Val Thr Ile Thr Ser Ala Asn Thr Gly Glu Leu Phe
1 5 10 15
<210> 95
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 95
Ala Thr Ser Val Gln Thr Gly Ala Leu Ile Tyr Glu Gln Tyr
1 5 10
<210> 96
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 96
Ala Ser Ser Pro Asp Arg Asp Arg Gly Asn Gln Pro Gln His
1 5 10
<210> 97
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 97
Ala Trp Lys Gly Arg Glu Tyr Glu Gln Tyr
1 5 10
<210> 98
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 98
Ser Ala Arg Gly Ser Gly Asp Asn Thr Gln Tyr
1 5 10
<210> 99
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 99
Ser Ala Pro Ala Ser Tyr Thr Glu Ala Phe
1 5 10
<210> 100
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 100
Ala Ser Ser Trp Ala Gly Leu Ser Tyr Glu Gln Tyr
1 5 10
Claims (7)
1.一种大肠癌的外周血TCR标志物,其特征在于,包括序列SpecSeq1-100所示蛋白中的至少一种。
2.根据权利要求1所述的大肠癌的外周血TCR标志物,其特征在于,所述标志物的蛋白序列为SpecSeq1-100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或2所述的标志物在制备检测或治疗大肠癌的制剂中的应用。
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细胞受体的质粒、病毒载体或核酸片段。
5.一种用于大肠癌检测的试剂盒,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。
7.一种检测大肠癌的蛋白质芯片,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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CN112521484A (zh) * | 2020-12-03 | 2021-03-19 | 佛山市第一人民医院(中山大学附属佛山医院) | 结肠癌肿瘤特异tcr序列及其应用 |
CN113030473A (zh) * | 2021-03-15 | 2021-06-25 | 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 |
WO2022012279A1 (zh) * | 2020-07-11 | 2022-01-20 | 成都益安博生物技术有限公司 | 一种大肠癌的外周血tcr标志物序列及其检测试剂盒和应用 |
WO2022194036A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种经典霍奇金淋巴瘤的外周血tcr标志物及其检测试剂盒和应用 |
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WO2022012279A1 (zh) * | 2020-07-11 | 2022-01-20 | 成都益安博生物技术有限公司 | 一种大肠癌的外周血tcr标志物序列及其检测试剂盒和应用 |
CN112521484A (zh) * | 2020-12-03 | 2021-03-19 | 佛山市第一人民医院(中山大学附属佛山医院) | 结肠癌肿瘤特异tcr序列及其应用 |
CN113030473A (zh) * | 2021-03-15 | 2021-06-25 | 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 |
WO2022194036A1 (zh) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | 一种经典霍奇金淋巴瘤的外周血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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