实施例6:CAR-F促进肿瘤类器官的形成和生长
肿瘤存在一个复杂的微环境系统,其中肿瘤中的成纤维细胞被称为CAFs(CancerAssocicated Fibroblasts),CAFs能促进肿瘤细胞的生长,转移以及在循环系统的生存能力。CAFs通过分泌生长因子、趋化因子和膜蛋白促进肿瘤细胞的生长、转移和浸润,这些因子包括白介素6(IL-6)、肝细胞生长因子(Hepatocyte Growth Factor,HGF)、成纤维细胞分泌蛋白1(Fibroblast Secreted Protein-1,FSP1)等(16;17)。在体外类器官模型中,加入工程化改造的成纤维细胞,可能促进肿瘤上皮细胞的自组装活力和提供更接近体内的生长环境,从而促进类器官的形成和生长。
在某些案例中,从非小细胞肺癌建立的PDX(Patient Derived Xenograft)模型或者从非小细胞肺癌外科手术中获得切除的肿瘤组织。蛋白酶酶解后,通过细胞过滤网,离心获取肿瘤上皮细胞。
肿瘤细胞分离合纯化的具体具体步骤如下:用蛋白酶稀释液制备得到蛋白酶复合工作液,50ul蛋白酶混合液(K2Oncology,#K2O-CRCEM-200L)加入9950ul蛋白酶稀释液(K2Oncology,#K2O-EDB)中。将组织块放入平皿中,加入4ml磷酸盐缓冲液清洗。吸出磷酸盐缓冲液,加入0.5ml,蛋白酶复合工作液。用剪刀将组织剪至匀浆状。加入4ml蛋白酶复合工作液,转移到24孔低贴壁细胞板中,放置于37℃孵育1h,显微镜下观察组织块酶解的程度。当观察到大量单细胞和寡细胞团时,用5ml移液管吸取组织匀浆,加入70微米尼龙网过滤到50ml离心管内,加入Advanced DMEM细胞培养液(Gibico,#12491-015),一并收集入细胞滤网。用5ml注射器的活塞,研磨滤网中的组织块,加入10-20ml细胞培养液A(#K2O-TWB),直到大部分组织通过滤网。离心200g,3分钟,加入30ml细胞培养液A(#K2O-TWB),再200g,3分钟离心收沉淀,用5-10ml细胞培养液A(#K2O-TWB)重悬。细胞计数和活力镜检,取10ul细胞悬液,加入10ul台盼蓝,观察活细胞比例,计算细胞密度。
在某些案例中,肿瘤上皮细胞和成纤维细胞,CTL-F(3T3-vehlcle)或者CAR-F细胞按照1:1比例混合,在基质胶中生长成肿瘤类器官。如图10中A所示,肿瘤细胞和对照组成纤维细胞混合形成3D肿瘤类器官培养结构,当表达不同嵌合抗原受体蛋白的成纤维细胞(CAR-Fs,此案例中为“3T3-EpHL-V”、“3T3-EpLH-V”、“3T3-EpHL-GP”、“3T3-EpLH-GP”、“3T3-EpHL-C8”、“3T3-EpLH-C8”)和肿瘤细胞按照1:1培养后,比对照组的成纤维细胞形成更多的肿瘤类器官细胞团,并且可以看到细胞生长速度更快。实验证明,CAR-F可以提高类器官模型培养的成功率和培养速度(图10,使用EpHL-V-510和EpLH-V-510,EpHL-C8-510和EpLH-C8-510,EpHL-GP-510和EpLH-GP-510)。
在某些案例中,原代肿瘤上皮细胞和工程化改造的成纤维细胞(CAR-F,此案例中为3T3-EpHL-C8)按照细胞数1:0.1、1:0.2、1:0.5、1:0.6、1:0.8、1:1、1:2、1:3、1:4、1:5混合基质胶进行3D肿瘤类器官培养。观察3D培养中,形成肿瘤类器官的数目和大小。实验证明,CAR-F可以提高类器官模型培养的成功率和培养速度,肿瘤细胞和CAR-F细胞混合比例在1:0.1~1:5之间。(图11,使用EpHL-V-510和EpLH-V-510,EpHL-C8-510和EpLH-C8-510,EpHL-GP-510和EpLH-GP-510)
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、改进等,均应包含在本发明的保护范围之内。
SEQUENCE LISTING
<110> 浙江科途医学科技有限公司
<120> 一种嵌合抗原受体成纤维细胞及其建立方法和应用
<130> CP11801134C
<160> 62
<170> PatentIn version 3.3
<210> 1
<211> 57
<212> DNA
<213> 信号肽编码核酸序列
<400> 1
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactcc 57
<210> 2
<211> 19
<212> PRT
<213> 信号肽氨基酸序列
<400> 2
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser
<210> 3
<211> 234
<212> DNA
<213> 包括跨膜序列的连接序列的编码核酸序列
<400> 3
agttctgtcg tgccagtcct tcagaaagtg aactctacta ctaccaagcc agtgctgcga 60
actccctcac ctgtgcaccc taccgggaca tctcagcccc agagaccaga agattgtcgg 120
ccccgtggct cagtgaaggg gaccggattg gacttcgcct gtgatattta catctgggca 180
cccttggccg gaatctgcgt ggcccttctg ctgtccttga tcatcactct catc 234
<210> 4
<211> 78
<212> PRT
<213> 包括跨膜序列的连接序列的氨基酸序列
<400> 4
Ser Ser Val Val Pro Val Leu Gln Lys Val Asn Ser Thr Thr Thr Lys
1 5 10 15
Pro Val Leu Arg Thr Pro Ser Pro Val His Pro Thr Gly Thr Ser Gln
20 25 30
Pro Gln Arg Pro Glu Asp Cys Arg Pro Arg Gly Ser Val Lys Gly Thr
35 40 45
Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly
50 55 60
Ile Cys Val Ala Leu Leu Leu Ser Leu Ile Ile Thr Leu Ile
65 70 75
<210> 5
<211> 90
<212> DNA
<213> 小鼠CD8细胞内序列的编码核酸序列
<400> 5
tgctaccaca ggagccgaaa gcgtgtttgc aaatgtccca ggccgctagt cagacaggaa 60
ggcaagccca gaccttcaga gaaaattgtg 90
<210> 6
<211> 30
<212> PRT
<213> 小鼠CD8细胞内序列的氨基酸序列
<400> 6
Cys Tyr His Arg Ser Arg Lys Arg Val Cys Lys Cys Pro Arg Pro Leu
1 5 10 15
Val Arg Gln Glu Gly Lys Pro Arg Pro Ser Glu Lys Ile Val
20 25 30
<210> 7
<211> 27
<212> DNA
<213> HA标签的编码核酸序列
<400> 7
tacccatacg atgttccaga ttacgct 27
<210> 8
<211> 9
<212> PRT
<213> HA标签的氨基酸序列
<400> 8
Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1 5
<210> 9
<211> 717
<212> DNA
<213> 绿色荧光蛋白标签的编码核酸序列
<400> 9
atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 60
ggcgacgtaa acggccacaa gttcagcgtg tctggcgagg gcgagggcga tgccacctac 120
ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 180
ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 240
cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 300
ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 360
gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 420
aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 480
ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 540
gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 600
tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 660
ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaag 717
<210> 10
<211> 239
<212> PRT
<213> 绿色荧光蛋白标签的氨基酸序列
<400> 10
Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu
1 5 10 15
Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly
20 25 30
Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile
35 40 45
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr
50 55 60
Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys
65 70 75 80
Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
85 90 95
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu
100 105 110
Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
115 120 125
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr
130 135 140
Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn
145 150 155 160
Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser
165 170 175
Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly
180 185 190
Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu
195 200 205
Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe
210 215 220
Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys
225 230 235
<210> 11
<211> 726
<212> DNA
<213> Anti-EpCAM ScfvVH-VL-31编码核酸序列
<400> 11
gctcgtcatg acccagtctc catcttatct tgctgcatct cctggagaaa ccattactat 60
taattgcagg gcaagtaaga gcattagcaa atatttagcc tggtatcaag agaaacctgg 120
gaaaactaat aagcttctta tctactctgg atccactttg caatctggaa ttccatcaag 180
gttcagtggc agtggatctg gtacagattt cactctcacc atcagtagcc tggagcctga 240
agattttgca atgtattact gtcaacagca taatgaatat ccgtacacgt tcggaggggg 300
gaccaagctt gagatcaaag gtggtggtgg ttctggcggc ggcggctccg gtggtggtgg 360
ttctgagagg tgcagctgct cgagcagtct ggagctgagc tggtgaaacc tggggcctca 420
gtgaagatat cctgcaaggc ttctggatac gccttcacta actactggct aggttgggta 480
aagcagaggc ctggacatgg acttgagtgg attggagatc ttttccctgg aagtggtaat 540
actcactaca atgagaggtt caggggcaaa gccacactga ctgcagacaa atcctcgagc 600
acagccttta tgcagctcag tagcctgaca tctgaggact ctgctgtcta tttctgtgca 660
agattgagga actgggacga ggctatggac tactggggcc aagggaccac ggtcaccgtc 720
tcctca 726
<210> 12
<211> 242
<212> PRT
<213> Anti-EpCAM ScfvVH-VL-31 氨基酸序列
<400> 12
Glu Leu Val Met Thr Gln Ser Pro Ser Tyr Leu Ala Ala Ser Pro Gly
1 5 10 15
Glu Thr Ile Thr Ile Asn Cys Arg Ala Ser Lys Ser Ile Ser Lys Tyr
20 25 30
Leu Ala Trp Tyr Gln Glu Lys Pro Gly Lys Thr Asn Lys Leu Leu Ile
35 40 45
Tyr Ser Gly Ser Thr Leu Gln Ser Gly Ile Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Met Tyr Tyr Cys Gln Gln His Asn Glu Tyr Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Leu Glu
115 120 125
Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys Ile Ser
130 135 140
Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Trp Leu Gly Trp Val
145 150 155 160
Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile Gly Asp Leu Phe Pro
165 170 175
Gly Ser Gly Asn Thr His Tyr Asn Glu Arg Phe Arg Gly Lys Ala Thr
180 185 190
Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Phe Met Gln Leu Ser Ser
195 200 205
Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Arg Asn
210 215 220
Trp Asp Glu Ala Met Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val
225 230 235 240
Ser Ser
<210> 13
<211> 726
<212> DNA
<213> Anti-EpCAM ScfvVL-VH-31的编码核酸序列
<400> 13
gaggtgcagc tgctcgagca gtctggagct gagctggtga aacctggggc ctcagtgaag 60
atatcctgca aggcttctgg atacgccttc actaactact ggctaggttg ggtaaagcag 120
aggcctggac atggacttga gtggattgga gatcttttcc ctggaagtgg taatactcac 180
tacaatgaga ggttcagggg caaagccaca ctgactgcag acaaatcctc gagcacagcc 240
tttatgcagc tcagtagcct gacatctgag gactctgctg tctatttctg tgcaagattg 300
aggaactggg acgaggctat ggactactgg ggccaaggga ccacggtcac cgtctcctca 360
ggtggtggtg gttctggcgg cggcggctcc ggtggtggtg gttctgagct cgtcatgacc 420
cagtctccat cttatcttgc tgcatctcct ggagaaacca ttactattaa ttgcagggca 480
agtaagagca ttagcaaata tttagcctgg tatcaagaga aacctgggaa aactaataag 540
cttcttatct actctggatc cactttgcaa tctggaattc catcaaggtt cagtggcagt 600
ggatctggta cagatttcac tctcaccatc agtagcctgg agcctgaaga ttttgcaatg 660
tattactgtc aacagcataa tgaatatccg tacacgttcg gaggggggac caagcttgag 720
atcaaa 726
<210> 14
<211> 242
<212> PRT
<213> Anti-EpCAM ScfvVL-VH-31氨基酸序列
<400> 14
Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val Lys Pro Gly
1 5 10 15
Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn
20 25 30
Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp
35 40 45
Ile Gly Asp Leu Phe Pro Gly Ser Gly Asn Thr His Tyr Asn Glu Arg
50 55 60
Phe Arg Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala
65 70 75 80
Phe Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe
85 90 95
Cys Ala Arg Leu Arg Asn Trp Asp Glu Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Glu Leu Val Met Thr Gln Ser Pro Ser
130 135 140
Tyr Leu Ala Ala Ser Pro Gly Glu Thr Ile Thr Ile Asn Cys Arg Ala
145 150 155 160
Ser Lys Ser Ile Ser Lys Tyr Leu Ala Trp Tyr Gln Glu Lys Pro Gly
165 170 175
Lys Thr Asn Lys Leu Leu Ile Tyr Ser Gly Ser Thr Leu Gln Ser Gly
180 185 190
Ile Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
195 200 205
Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Met Tyr Tyr Cys Gln
210 215 220
Gln His Asn Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Lys
<210> 15
<211> 620
<212> DNA
<213> Anti-EpCAM ScfvVH-VL-47编码核酸序列
<400> 15
gctcgtgatg acccagactc cactctccct gcctgtcagt cttggagatc aagcctccat 60
ctcttgcaga tctagtcaga gccttgtaca cagtaatgga aacacctatt tacattggta 120
cctgcagaag ccaggccagt ctccaaagct cctgatctac aaagtttcca accgattttc 180
tggggtccca gacaggttca gtggcagtgg atcagggaca gatttcacac tcaagatcag 240
cagagtggag gctgaggatc tgggagttta tttctgctct caaagtacac atgttccgta 300
cacgttcgga ggggggacca agcttgagat caaaggtggt ggtggttctg gcggcggcgg 360
ctccggtggt ggtggttctg atccttgagt ggattggaga ggtttatcct agaattggta 420
atgcttacta caatgagaag ttcaagggca aggccacact gactgcagac aaatcctcca 480
gcacagcgtc catggagctc cgcagcctga cctctgagga ctctgcggtc tatttctgtg 540
caagacgggg atcctacgat actaactacg actggtactt cgatgtctgg ggccaaggga 600
ccacggtcac cgtctcctca 620
<210> 16
<211> 251
<212> PRT
<213> Anti-EpCAM ScfvVH-VL-47氨基酸序列
<400> 16
Glu Leu Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser
85 90 95
Thr His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu
115 120 125
Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala
130 135 140
Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr
145 150 155 160
Gly Leu Ser Trp Val Lys Gln Arg Pro Gly Gln Val Leu Glu Trp Ile
165 170 175
Gly Glu Val Tyr Pro Arg Ile Gly Asn Ala Tyr Tyr Asn Glu Lys Phe
180 185 190
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Ser
195 200 205
Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys
210 215 220
Ala Arg Arg Gly Ser Tyr Asp Thr Asn Tyr Asp Trp Tyr Phe Asp Val
225 230 235 240
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
245 250
<210> 17
<211> 620
<212> DNA
<213> Anti-EpCAM ScfvVL-VH-47编码核酸序列
<400> 17
tccttgagtg gattggagag gtttatccta gaattggtaa tgcttactac aatgagaagt 60
tcaagggcaa ggccacactg actgcagaca aatcctccag cacagcgtcc atggagctcc 120
gcagcctgac ctctgaggac tctgcggtct atttctgtgc aagacgggga tcctacgata 180
ctaactacga ctggtacttc gatgtctggg gccaagggac cacggtcacc gtctcctcag 240
gtggtggtgg ttctggcggc ggcggctccg gtggtggtgg ttctgagctc gtgatgaccc 300
agactccact ctccctgcct gtcagtcttg gagatcaagc ctccatctct tgcagatcta 360
gtcagagcct tgtacacagt aatggaaaca cctatttaca ttggtacctg cagaagccag 420
gccagtctcc aaagctcctg atctacaaag tttccaaccg attttctggg gtcccagaca 480
ggttcagtgg cagtggatca gggacagatt tcacactcaa gatcagcaga gtggaggctg 540
aggatctggg agtttatttc tgctctcaaa gtacacatgt tccgtacacg ttcggagggg 600
ggaccaagct tgagatcaaa 620
<210> 18
<211> 251
<212> PRT
<213> Anti-EpCAM ScfvVL-VH-47 氨基酸序列
<400> 18
Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly
1 5 10 15
Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn
20 25 30
Tyr Gly Leu Ser Trp Val Lys Gln Arg Pro Gly Gln Val Leu Glu Trp
35 40 45
Ile Gly Glu Val Tyr Pro Arg Ile Gly Asn Ala Tyr Tyr Asn Glu Lys
50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala
65 70 75 80
Ser Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe
85 90 95
Cys Ala Arg Arg Gly Ser Tyr Asp Thr Asn Tyr Asp Trp Tyr Phe Asp
100 105 110
Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly
115 120 125
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Val Met Thr
130 135 140
Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly Asp Gln Ala Ser Ile
145 150 155 160
Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr
165 170 175
Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile
180 185 190
Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly
195 200 205
Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala
210 215 220
Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser Thr His Val Pro Tyr
225 230 235 240
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
245 250
<210> 19
<211> 738
<212> DNA
<213> Anti-EpCAM ScfvVH-VL-510的编码核酸序列
<400> 19
gaggtgcagc tgctcgagca gtctggagct gagctggtaa ggcctgggac ttcagtgaag 60
atatcctgca aggcttctgg atacgccttc actaactact ggctaggttg ggtaaagcag 120
aggcctggac atggacttga gtggattgga gatattttcc ctggaagtgg taatatccac 180
tacaatgaga agttcaaggg caaagccaca ctgactgcag acaaatcttc gagcacagcc 240
tatatgcagc tcagtagcct gacatttgag gactctgctg tctatttctg tgcaagactg 300
aggaactggg acgagcctat ggactactgg ggccaaggga ccacggtcac cgtcggtggt 360
ggtggttctg gcggcggcgg ctccggtggt ggtggttctg agctcgtgat gacacagtct 420
ccatcctccc tgactgtgac agcaggagag aaggtcacta tgagctgcaa gtccagtcag 480
agtctgttaa acagtggaaa tcaaaagaac tacttgacct ggtaccagca gaaaccaggg 540
cagcctccta aactgttgat ctactgggca tccactaggg aatctggggt ccctgatcgc 600
ttcacaggca gtggatctgg aacagatttc actctcacca tcagcagtgt gcaggctgaa 660
gacctggcag tttattactg tcagaatgat tatagttatc cgctcacgtt cggtgctggg 720
accaagcttg agatcaaa 738
<210> 20
<211> 246
<212> PRT
<213> Anti-EpCAM ScfvVH-VL-510的氨基酸序列
<400> 20
Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val Arg Pro Gly
1 5 10 15
Thr Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn
20 25 30
Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp
35 40 45
Ile Gly Asp Ile Phe Pro Gly Ser Gly Asn Ile His Tyr Asn Glu Lys
50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala
65 70 75 80
Tyr Met Gln Leu Ser Ser Leu Thr Phe Glu Asp Ser Ala Val Tyr Phe
85 90 95
Cys Ala Arg Leu Arg Asn Trp Asp Glu Pro Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Thr Val Thr Val Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Leu Val Met Thr Gln Ser Pro Ser Ser Leu
130 135 140
Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln
145 150 155 160
Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln
165 170 175
Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr
180 185 190
Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr
195 200 205
Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val
210 215 220
Tyr Tyr Cys Gln Asn Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly
225 230 235 240
Thr Lys Leu Glu Ile Lys
245
<210> 21
<211> 738
<212> DNA
<213> Anti-EpCAM ScfvVL-VH-510的编码核酸序列
<400> 21
gagctcgtga tgacacagtc tccatcctcc ctgactgtga cagcaggaga gaaggtcact 60
atgagctgca agtccagtca gagtctgtta aacagtggaa atcaaaagaa ctacttgacc 120
tggtaccagc agaaaccagg gcagcctcct aaactgttga tctactgggc atccactagg 180
gaatctgggg tccctgatcg cttcacaggc agtggatctg gaacagattt cactctcacc 240
atcagcagtg tgcaggctga agacctggca gtttattact gtcagaatga ttatagttat 300
ccgctcacgt tcggtgctgg gaccaagctt gagatcaaag gtggtggtgg ttctggcggc 360
ggcggctccg gtggtggtgg ttctgaggtg cagctgctcg agcagtctgg agctgagctg 420
gtaaggcctg ggacttcagt gaagatatcc tgcaaggctt ctggatacgc cttcactaac 480
tactggctag gttgggtaaa gcagaggcct ggacatggac ttgagtggat tggagatatt 540
ttccctggaa gtggtaatat ccactacaat gagaagttca agggcaaagc cacactgact 600
gcagacaaat cttcgagcac agcctatatg cagctcagta gcctgacatt tgaggactct 660
gctgtctatt tctgtgcaag actgaggaac tgggacgagc ctatggacta ctggggccaa 720
gggaccacgg tcaccgtc 738
<210> 22
<211> 246
<212> PRT
<213> Anti-EpCAM ScfvVL-VH-510的氨基酸序列
<400> 22
Glu Leu Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly
1 5 10 15
Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30
Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Ile
100 105 110
Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val Arg Pro Gly
130 135 140
Thr Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn
145 150 155 160
Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp
165 170 175
Ile Gly Asp Ile Phe Pro Gly Ser Gly Asn Ile His Tyr Asn Glu Lys
180 185 190
Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala
195 200 205
Tyr Met Gln Leu Ser Ser Leu Thr Phe Glu Asp Ser Ala Val Tyr Phe
210 215 220
Cys Ala Arg Leu Arg Asn Trp Asp Glu Pro Met Asp Tyr Trp Gly Gln
225 230 235 240
Gly Thr Thr Val Thr Val
245
<210> 23
<211> 1047
<212> DNA
<213> 嵌合抗原受体蛋白EpHL-V-31编码核酸序列
<400> 23
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgct 60
cgtcatgacc cagtctccat cttatcttgc tgcatctcct ggagaaacca ttactattaa 120
ttgcagggca agtaagagca ttagcaaata tttagcctgg tatcaagaga aacctgggaa 180
aactaataag cttcttatct actctggatc cactttgcaa tctggaattc catcaaggtt 240
cagtggcagt ggatctggta cagatttcac tctcaccatc agtagcctgg agcctgaaga 300
ttttgcaatg tattactgtc aacagcataa tgaatatccg tacacgttcg gaggggggac 360
caagcttgag atcaaaggtg gtggtggttc tggcggcggc ggctccggtg gtggtggttc 420
tgagaggtgc agctgctcga gcagtctgga gctgagctgg tgaaacctgg ggcctcagtg 480
aagatatcct gcaaggcttc tggatacgcc ttcactaact actggctagg ttgggtaaag 540
cagaggcctg gacatggact tgagtggatt ggagatcttt tccctggaag tggtaatact 600
cactacaatg agaggttcag gggcaaagcc acactgactg cagacaaatc ctcgagcaca 660
gcctttatgc agctcagtag cctgacatct gaggactctg ctgtctattt ctgtgcaaga 720
ttgaggaact gggacgaggc tatggactac tggggccaag ggaccacggt caccgtctcc 780
tcaagttctg tcgtgccagt ccttcagaaa gtgaactcta ctactaccaa gccagtgctg 840
cgaactccct cacctgtgca ccctaccggg acatctcagc cccagagacc agaagattgt 900
cggccccgtg gctcagtgaa ggggaccgga ttggacttcg cctgtgatat ttacatctgg 960
gcacccttgg ccggaatctg cgtggccctt ctgctgtcct tgatcatcac tctcatctac 1020
ccatacgatg ttccagatta cgcttaa 1047
<210> 24
<211> 348
<212> PRT
<213> 嵌合抗原受体蛋白EpHL-V-31氨基酸序列
<400> 24
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Leu Val Met Thr Gln Ser Pro Ser Tyr Leu Ala Ala
20 25 30
Ser Pro Gly Glu Thr Ile Thr Ile Asn Cys Arg Ala Ser Lys Ser Ile
35 40 45
Ser Lys Tyr Leu Ala Trp Tyr Gln Glu Lys Pro Gly Lys Thr Asn Lys
50 55 60
Leu Leu Ile Tyr Ser Gly Ser Thr Leu Gln Ser Gly Ile Pro Ser Arg
65 70 75 80
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
85 90 95
Leu Glu Pro Glu Asp Phe Ala Met Tyr Tyr Cys Gln Gln His Asn Glu
100 105 110
Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln
130 135 140
Leu Leu Glu Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val
145 150 155 160
Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Trp Leu
165 170 175
Gly Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile Gly Asp
180 185 190
Leu Phe Pro Gly Ser Gly Asn Thr His Tyr Asn Glu Arg Phe Arg Gly
195 200 205
Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Phe Met Gln
210 215 220
Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg
225 230 235 240
Leu Arg Asn Trp Asp Glu Ala Met Asp Tyr Trp Gly Gln Gly Thr Thr
245 250 255
Val Thr Val Ser Ser Ser Ser Val Val Pro Val Leu Gln Lys Val Asn
260 265 270
Ser Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser Pro Val His Pro
275 280 285
Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys Arg Pro Arg Gly
290 295 300
Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp
305 310 315 320
Ala Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu Ser Leu Ile Ile
325 330 335
Thr Leu Ile Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
340 345
<210> 25
<211> 1047
<212> DNA
<213> 嵌合抗原受体蛋白EpLH-V-31编码核酸序列
<400> 25
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgag 60
gtgcagctgc tcgagcagtc tggagctgag ctggtgaaac ctggggcctc agtgaagata 120
tcctgcaagg cttctggata cgccttcact aactactggc taggttgggt aaagcagagg 180
cctggacatg gacttgagtg gattggagat cttttccctg gaagtggtaa tactcactac 240
aatgagaggt tcaggggcaa agccacactg actgcagaca aatcctcgag cacagccttt 300
atgcagctca gtagcctgac atctgaggac tctgctgtct atttctgtgc aagattgagg 360
aactgggacg aggctatgga ctactggggc caagggacca cggtcaccgt ctcctcaggt 420
ggtggtggtt ctggcggcgg cggctccggt ggtggtggtt ctgagctcgt catgacccag 480
tctccatctt atcttgctgc atctcctgga gaaaccatta ctattaattg cagggcaagt 540
aagagcatta gcaaatattt agcctggtat caagagaaac ctgggaaaac taataagctt 600
cttatctact ctggatccac tttgcaatct ggaattccat caaggttcag tggcagtgga 660
tctggtacag atttcactct caccatcagt agcctggagc ctgaagattt tgcaatgtat 720
tactgtcaac agcataatga atatccgtac acgttcggag gggggaccaa gcttgagatc 780
aaaagttctg tcgtgccagt ccttcagaaa gtgaactcta ctactaccaa gccagtgctg 840
cgaactccct cacctgtgca ccctaccggg acatctcagc cccagagacc agaagattgt 900
cggccccgtg gctcagtgaa ggggaccgga ttggacttcg cctgtgatat ttacatctgg 960
gcacccttgg ccggaatctg cgtggccctt ctgctgtcct tgatcatcac tctcatctac 1020
ccatacgatg ttccagatta cgcttaa 1047
<210> 26
<211> 348
<212> PRT
<213> 嵌合抗原受体蛋白EpLH-V-31氨基酸序列
<400> 26
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val
20 25 30
Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala
35 40 45
Phe Thr Asn Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly
50 55 60
Leu Glu Trp Ile Gly Asp Leu Phe Pro Gly Ser Gly Asn Thr His Tyr
65 70 75 80
Asn Glu Arg Phe Arg Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Phe Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala
100 105 110
Val Tyr Phe Cys Ala Arg Leu Arg Asn Trp Asp Glu Ala Met Asp Tyr
115 120 125
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser
130 135 140
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Val Met Thr Gln
145 150 155 160
Ser Pro Ser Tyr Leu Ala Ala Ser Pro Gly Glu Thr Ile Thr Ile Asn
165 170 175
Cys Arg Ala Ser Lys Ser Ile Ser Lys Tyr Leu Ala Trp Tyr Gln Glu
180 185 190
Lys Pro Gly Lys Thr Asn Lys Leu Leu Ile Tyr Ser Gly Ser Thr Leu
195 200 205
Gln Ser Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
210 215 220
Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Met Tyr
225 230 235 240
Tyr Cys Gln Gln His Asn Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr
245 250 255
Lys Leu Glu Ile Lys Ser Ser Val Val Pro Val Leu Gln Lys Val Asn
260 265 270
Ser Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser Pro Val His Pro
275 280 285
Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys Arg Pro Arg Gly
290 295 300
Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp
305 310 315 320
Ala Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu Ser Leu Ile Ile
325 330 335
Thr Leu Ile Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
340 345
<210> 27
<211> 941
<212> DNA
<213> 嵌合抗原受体蛋白EpHL-V-47编码核酸序列
<400> 27
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgct 60
cgtgatgacc cagactccac tctccctgcc tgtcagtctt ggagatcaag cctccatctc 120
ttgcagatct agtcagagcc ttgtacacag taatggaaac acctatttac attggtacct 180
gcagaagcca ggccagtctc caaagctcct gatctacaaa gtttccaacc gattttctgg 240
ggtcccagac aggttcagtg gcagtggatc agggacagat ttcacactca agatcagcag 300
agtggaggct gaggatctgg gagtttattt ctgctctcaa agtacacatg ttccgtacac 360
gttcggaggg gggaccaagc ttgagatcaa aggtggtggt ggttctggcg gcggcggctc 420
cggtggtggt ggttctgatc cttgagtgga ttggagaggt ttatcctaga attggtaatg 480
cttactacaa tgagaagttc aagggcaagg ccacactgac tgcagacaaa tcctccagca 540
cagcgtccat ggagctccgc agcctgacct ctgaggactc tgcggtctat ttctgtgcaa 600
gacggggatc ctacgatact aactacgact ggtacttcga tgtctggggc caagggacca 660
cggtcaccgt ctcctcaagt tctgtcgtgc cagtccttca gaaagtgaac tctactacta 720
ccaagccagt gctgcgaact ccctcacctg tgcaccctac cgggacatct cagccccaga 780
gaccagaaga ttgtcggccc cgtggctcag tgaaggggac cggattggac ttcgcctgtg 840
atatttacat ctgggcaccc ttggccggaa tctgcgtggc ccttctgctg tccttgatca 900
tcactctcat ctacccatac gatgttccag attacgctta a 941
<210> 28
<211> 357
<212> PRT
<213> 嵌合抗原受体蛋白EpHL-V-47氨基酸序列
<400> 28
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Leu Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val
20 25 30
Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu
35 40 45
Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro
50 55 60
Gly Gln Ser Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser
65 70 75 80
Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
85 90 95
Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys
100 105 110
Ser Gln Ser Thr His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu
115 120 125
Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Ala Arg
145 150 155 160
Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe
165 170 175
Thr Asn Tyr Gly Leu Ser Trp Val Lys Gln Arg Pro Gly Gln Val Leu
180 185 190
Glu Trp Ile Gly Glu Val Tyr Pro Arg Ile Gly Asn Ala Tyr Tyr Asn
195 200 205
Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser
210 215 220
Thr Ala Ser Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val
225 230 235 240
Tyr Phe Cys Ala Arg Arg Gly Ser Tyr Asp Thr Asn Tyr Asp Trp Tyr
245 250 255
Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ser Ser
260 265 270
Val Val Pro Val Leu Gln Lys Val Asn Ser Thr Thr Thr Lys Pro Val
275 280 285
Leu Arg Thr Pro Ser Pro Val His Pro Thr Gly Thr Ser Gln Pro Gln
290 295 300
Arg Pro Glu Asp Cys Arg Pro Arg Gly Ser Val Lys Gly Thr Gly Leu
305 310 315 320
Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Ile Cys
325 330 335
Val Ala Leu Leu Leu Ser Leu Ile Ile Thr Leu Ile Tyr Pro Tyr Asp
340 345 350
Val Pro Asp Tyr Ala
355
<210> 29
<211> 941
<212> DNA
<213> 嵌合抗原受体蛋白EpLH-V-47编码核酸序列
<400> 29
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactcctcc 60
ttgagtggat tggagaggtt tatcctagaa ttggtaatgc ttactacaat gagaagttca 120
agggcaaggc cacactgact gcagacaaat cctccagcac agcgtccatg gagctccgca 180
gcctgacctc tgaggactct gcggtctatt tctgtgcaag acggggatcc tacgatacta 240
actacgactg gtacttcgat gtctggggcc aagggaccac ggtcaccgtc tcctcaggtg 300
gtggtggttc tggcggcggc ggctccggtg gtggtggttc tgagctcgtg atgacccaga 360
ctccactctc cctgcctgtc agtcttggag atcaagcctc catctcttgc agatctagtc 420
agagccttgt acacagtaat ggaaacacct atttacattg gtacctgcag aagccaggcc 480
agtctccaaa gctcctgatc tacaaagttt ccaaccgatt ttctggggtc ccagacaggt 540
tcagtggcag tggatcaggg acagatttca cactcaagat cagcagagtg gaggctgagg 600
atctgggagt ttatttctgc tctcaaagta cacatgttcc gtacacgttc ggagggggga 660
ccaagcttga gatcaaaagt tctgtcgtgc cagtccttca gaaagtgaac tctactacta 720
ccaagccagt gctgcgaact ccctcacctg tgcaccctac cgggacatct cagccccaga 780
gaccagaaga ttgtcggccc cgtggctcag tgaaggggac cggattggac ttcgcctgtg 840
atatttacat ctgggcaccc ttggccggaa tctgcgtggc ccttctgctg tccttgatca 900
tcactctcat ctacccatac gatgttccag attacgctta a 941
<210> 30
<211> 357
<212> PRT
<213> 嵌合抗原受体蛋白EpLH-V-47氨基酸序列
<400> 30
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Ala
20 25 30
Arg Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr
35 40 45
Phe Thr Asn Tyr Gly Leu Ser Trp Val Lys Gln Arg Pro Gly Gln Val
50 55 60
Leu Glu Trp Ile Gly Glu Val Tyr Pro Arg Ile Gly Asn Ala Tyr Tyr
65 70 75 80
Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Ser Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala
100 105 110
Val Tyr Phe Cys Ala Arg Arg Gly Ser Tyr Asp Thr Asn Tyr Asp Trp
115 120 125
Tyr Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly
130 135 140
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu
145 150 155 160
Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly Asp Gln
165 170 175
Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly
180 185 190
Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Lys
195 200 205
Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp Arg
210 215 220
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg
225 230 235 240
Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser Thr His
245 250 255
Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Ser Ser
260 265 270
Val Val Pro Val Leu Gln Lys Val Asn Ser Thr Thr Thr Lys Pro Val
275 280 285
Leu Arg Thr Pro Ser Pro Val His Pro Thr Gly Thr Ser Gln Pro Gln
290 295 300
Arg Pro Glu Asp Cys Arg Pro Arg Gly Ser Val Lys Gly Thr Gly Leu
305 310 315 320
Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Ile Cys
325 330 335
Val Ala Leu Leu Leu Ser Leu Ile Ile Thr Leu Ile Tyr Pro Tyr Asp
340 345 350
Val Pro Asp Tyr Ala
355
<210> 31
<211> 1059
<212> DNA
<213> 嵌合抗原受体蛋白EpHL-V-510编码核酸序列
<400> 31
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgag 60
gtgcagctgc tcgagcagtc tggagctgag ctggtaaggc ctgggacttc agtgaagata 120
tcctgcaagg cttctggata cgccttcact aactactggc taggttgggt aaagcagagg 180
cctggacatg gacttgagtg gattggagat attttccctg gaagtggtaa tatccactac 240
aatgagaagt tcaagggcaa agccacactg actgcagaca aatcttcgag cacagcctat 300
atgcagctca gtagcctgac atttgaggac tctgctgtct atttctgtgc aagactgagg 360
aactgggacg agcctatgga ctactggggc caagggacca cggtcaccgt cggtggtggt 420
ggttctggcg gcggcggctc cggtggtggt ggttctgagc tcgtgatgac acagtctcca 480
tcctccctga ctgtgacagc aggagagaag gtcactatga gctgcaagtc cagtcagagt 540
ctgttaaaca gtggaaatca aaagaactac ttgacctggt accagcagaa accagggcag 600
cctcctaaac tgttgatcta ctgggcatcc actagggaat ctggggtccc tgatcgcttc 660
acaggcagtg gatctggaac agatttcact ctcaccatca gcagtgtgca ggctgaagac 720
ctggcagttt attactgtca gaatgattat agttatccgc tcacgttcgg tgctgggacc 780
aagcttgaga tcaaaagttc tgtcgtgcca gtccttcaga aagtgaactc tactactacc 840
aagccagtgc tgcgaactcc ctcacctgtg caccctaccg ggacatctca gccccagaga 900
ccagaagatt gtcggccccg tggctcagtg aaggggaccg gattggactt cgcctgtgat 960
atttacatct gggcaccctt ggccggaatc tgcgtggccc ttctgctgtc cttgatcatc 1020
actctcatct acccatacga tgttccagat tacgcttaa 1059
<210> 32
<211> 352
<212> PRT
<213> 嵌合抗原受体蛋白EpHL-V-510氨基酸序列
<400> 32
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val
20 25 30
Arg Pro Gly Thr Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala
35 40 45
Phe Thr Asn Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly
50 55 60
Leu Glu Trp Ile Gly Asp Ile Phe Pro Gly Ser Gly Asn Ile His Tyr
65 70 75 80
Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Phe Glu Asp Ser Ala
100 105 110
Val Tyr Phe Cys Ala Arg Leu Arg Asn Trp Asp Glu Pro Met Asp Tyr
115 120 125
Trp Gly Gln Gly Thr Thr Val Thr Val Gly Gly Gly Gly Ser Gly Gly
130 135 140
Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Val Met Thr Gln Ser Pro
145 150 155 160
Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys
165 170 175
Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu Thr
180 185 190
Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp
195 200 205
Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly
210 215 220
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp
225 230 235 240
Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Ser Tyr Pro Leu Thr Phe
245 250 255
Gly Ala Gly Thr Lys Leu Glu Ile Lys Ser Ser Val Val Pro Val Leu
260 265 270
Gln Lys Val Asn Ser Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser
275 280 285
Pro Val His Pro Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys
290 295 300
Arg Pro Arg Gly Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp
305 310 315 320
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu
325 330 335
Ser Leu Ile Ile Thr Leu Ile Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
340 345 350
<210> 33
<211> 1059
<212> DNA
<213> 嵌合抗原受体蛋白EpLH-V-510编码核酸序列
<400> 33
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgag 60
ctcgtgatga cacagtctcc atcctccctg actgtgacag caggagagaa ggtcactatg 120
agctgcaagt ccagtcagag tctgttaaac agtggaaatc aaaagaacta cttgacctgg 180
taccagcaga aaccagggca gcctcctaaa ctgttgatct actgggcatc cactagggaa 240
tctggggtcc ctgatcgctt cacaggcagt ggatctggaa cagatttcac tctcaccatc 300
agcagtgtgc aggctgaaga cctggcagtt tattactgtc agaatgatta tagttatccg 360
ctcacgttcg gtgctgggac caagcttgag atcaaaggtg gtggtggttc tggcggcggc 420
ggctccggtg gtggtggttc tgaggtgcag ctgctcgagc agtctggagc tgagctggta 480
aggcctggga cttcagtgaa gatatcctgc aaggcttctg gatacgcctt cactaactac 540
tggctaggtt gggtaaagca gaggcctgga catggacttg agtggattgg agatattttc 600
cctggaagtg gtaatatcca ctacaatgag aagttcaagg gcaaagccac actgactgca 660
gacaaatctt cgagcacagc ctatatgcag ctcagtagcc tgacatttga ggactctgct 720
gtctatttct gtgcaagact gaggaactgg gacgagccta tggactactg gggccaaggg 780
accacggtca ccgtcagttc tgtcgtgcca gtccttcaga aagtgaactc tactactacc 840
aagccagtgc tgcgaactcc ctcacctgtg caccctaccg ggacatctca gccccagaga 900
ccagaagatt gtcggccccg tggctcagtg aaggggaccg gattggactt cgcctgtgat 960
atttacatct gggcaccctt ggccggaatc tgcgtggccc ttctgctgtc cttgatcatc 1020
actctcatct acccatacga tgttccagat tacgcttaa 1059
<210> 34
<211> 352
<212> PRT
<213> 嵌合抗原受体蛋白EpLH-V-510氨基酸序列
<400> 34
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Leu Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val
20 25 30
Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu
35 40 45
Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys
50 55 60
Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe
85 90 95
Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr
100 105 110
Cys Gln Asn Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys
115 120 125
Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
130 135 140
Gly Gly Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val
145 150 155 160
Arg Pro Gly Thr Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala
165 170 175
Phe Thr Asn Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly
180 185 190
Leu Glu Trp Ile Gly Asp Ile Phe Pro Gly Ser Gly Asn Ile His Tyr
195 200 205
Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
210 215 220
Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Phe Glu Asp Ser Ala
225 230 235 240
Val Tyr Phe Cys Ala Arg Leu Arg Asn Trp Asp Glu Pro Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Val Val Pro Val Leu
260 265 270
Gln Lys Val Asn Ser Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser
275 280 285
Pro Val His Pro Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys
290 295 300
Arg Pro Arg Gly Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp
305 310 315 320
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu
325 330 335
Ser Leu Ile Ile Thr Leu Ile Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
340 345 350
<210> 35
<211> 1137
<212> DNA
<213> 嵌合抗原受体蛋白EpHL-C8-31编码核酸序列
<400> 35
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgct 60
cgtcatgacc cagtctccat cttatcttgc tgcatctcct ggagaaacca ttactattaa 120
ttgcagggca agtaagagca ttagcaaata tttagcctgg tatcaagaga aacctgggaa 180
aactaataag cttcttatct actctggatc cactttgcaa tctggaattc catcaaggtt 240
cagtggcagt ggatctggta cagatttcac tctcaccatc agtagcctgg agcctgaaga 300
ttttgcaatg tattactgtc aacagcataa tgaatatccg tacacgttcg gaggggggac 360
caagcttgag atcaaaggtg gtggtggttc tggcggcggc ggctccggtg gtggtggttc 420
tgagaggtgc agctgctcga gcagtctgga gctgagctgg tgaaacctgg ggcctcagtg 480
aagatatcct gcaaggcttc tggatacgcc ttcactaact actggctagg ttgggtaaag 540
cagaggcctg gacatggact tgagtggatt ggagatcttt tccctggaag tggtaatact 600
cactacaatg agaggttcag gggcaaagcc acactgactg cagacaaatc ctcgagcaca 660
gcctttatgc agctcagtag cctgacatct gaggactctg ctgtctattt ctgtgcaaga 720
ttgaggaact gggacgaggc tatggactac tggggccaag ggaccacggt caccgtctcc 780
tcaagttctg tcgtgccagt ccttcagaaa gtgaactcta ctactaccaa gccagtgctg 840
cgaactccct cacctgtgca ccctaccggg acatctcagc cccagagacc agaagattgt 900
cggccccgtg gctcagtgaa ggggaccgga ttggacttcg cctgtgatat ttacatctgg 960
gcacccttgg ccggaatctg cgtggccctt ctgctgtcct tgatcatcac tctcatctgc 1020
taccacagga gccgaaagcg tgtttgcaaa tgtcccaggc cgctagtcag acaggaaggc 1080
aagcccagac cttcagagaa aattgtgtac ccatacgatg ttccagatta cgcttaa 1137
<210> 36
<211> 378
<212> PRT
<213> 嵌合抗原受体蛋白EpHL-C8-31氨基酸序列
<400> 36
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Leu Val Met Thr Gln Ser Pro Ser Tyr Leu Ala Ala
20 25 30
Ser Pro Gly Glu Thr Ile Thr Ile Asn Cys Arg Ala Ser Lys Ser Ile
35 40 45
Ser Lys Tyr Leu Ala Trp Tyr Gln Glu Lys Pro Gly Lys Thr Asn Lys
50 55 60
Leu Leu Ile Tyr Ser Gly Ser Thr Leu Gln Ser Gly Ile Pro Ser Arg
65 70 75 80
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
85 90 95
Leu Glu Pro Glu Asp Phe Ala Met Tyr Tyr Cys Gln Gln His Asn Glu
100 105 110
Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln
130 135 140
Leu Leu Glu Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val
145 150 155 160
Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Trp Leu
165 170 175
Gly Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile Gly Asp
180 185 190
Leu Phe Pro Gly Ser Gly Asn Thr His Tyr Asn Glu Arg Phe Arg Gly
195 200 205
Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Phe Met Gln
210 215 220
Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg
225 230 235 240
Leu Arg Asn Trp Asp Glu Ala Met Asp Tyr Trp Gly Gln Gly Thr Thr
245 250 255
Val Thr Val Ser Ser Ser Ser Val Val Pro Val Leu Gln Lys Val Asn
260 265 270
Ser Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser Pro Val His Pro
275 280 285
Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys Arg Pro Arg Gly
290 295 300
Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp
305 310 315 320
Ala Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu Ser Leu Ile Ile
325 330 335
Thr Leu Ile Cys Tyr His Arg Ser Arg Lys Arg Val Cys Lys Cys Pro
340 345 350
Arg Pro Leu Val Arg Gln Glu Gly Lys Pro Arg Pro Ser Glu Lys Ile
355 360 365
Val Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
370 375
<210> 37
<211> 1137
<212> DNA
<213> 嵌合抗原受体蛋白EpLH-C8-31编码核酸序列
<400> 37
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgag 60
gtgcagctgc tcgagcagtc tggagctgag ctggtgaaac ctggggcctc agtgaagata 120
tcctgcaagg cttctggata cgccttcact aactactggc taggttgggt aaagcagagg 180
cctggacatg gacttgagtg gattggagat cttttccctg gaagtggtaa tactcactac 240
aatgagaggt tcaggggcaa agccacactg actgcagaca aatcctcgag cacagccttt 300
atgcagctca gtagcctgac atctgaggac tctgctgtct atttctgtgc aagattgagg 360
aactgggacg aggctatgga ctactggggc caagggacca cggtcaccgt ctcctcaggt 420
ggtggtggtt ctggcggcgg cggctccggt ggtggtggtt ctgagctcgt catgacccag 480
tctccatctt atcttgctgc atctcctgga gaaaccatta ctattaattg cagggcaagt 540
aagagcatta gcaaatattt agcctggtat caagagaaac ctgggaaaac taataagctt 600
cttatctact ctggatccac tttgcaatct ggaattccat caaggttcag tggcagtgga 660
tctggtacag atttcactct caccatcagt agcctggagc ctgaagattt tgcaatgtat 720
tactgtcaac agcataatga atatccgtac acgttcggag gggggaccaa gcttgagatc 780
aaaagttctg tcgtgccagt ccttcagaaa gtgaactcta ctactaccaa gccagtgctg 840
cgaactccct cacctgtgca ccctaccggg acatctcagc cccagagacc agaagattgt 900
cggccccgtg gctcagtgaa ggggaccgga ttggacttcg cctgtgatat ttacatctgg 960
gcacccttgg ccggaatctg cgtggccctt ctgctgtcct tgatcatcac tctcatctgc 1020
taccacagga gccgaaagcg tgtttgcaaa tgtcccaggc cgctagtcag acaggaaggc 1080
aagcccagac cttcagagaa aattgtgtac ccatacgatg ttccagatta cgcttaa 1137
<210> 38
<211> 378
<212> PRT
<213> 嵌合抗原受体蛋白EpLH-C8-31氨基酸序列
<400> 38
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val
20 25 30
Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala
35 40 45
Phe Thr Asn Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly
50 55 60
Leu Glu Trp Ile Gly Asp Leu Phe Pro Gly Ser Gly Asn Thr His Tyr
65 70 75 80
Asn Glu Arg Phe Arg Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Phe Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala
100 105 110
Val Tyr Phe Cys Ala Arg Leu Arg Asn Trp Asp Glu Ala Met Asp Tyr
115 120 125
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser
130 135 140
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Val Met Thr Gln
145 150 155 160
Ser Pro Ser Tyr Leu Ala Ala Ser Pro Gly Glu Thr Ile Thr Ile Asn
165 170 175
Cys Arg Ala Ser Lys Ser Ile Ser Lys Tyr Leu Ala Trp Tyr Gln Glu
180 185 190
Lys Pro Gly Lys Thr Asn Lys Leu Leu Ile Tyr Ser Gly Ser Thr Leu
195 200 205
Gln Ser Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
210 215 220
Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Met Tyr
225 230 235 240
Tyr Cys Gln Gln His Asn Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr
245 250 255
Lys Leu Glu Ile Lys Ser Ser Val Val Pro Val Leu Gln Lys Val Asn
260 265 270
Ser Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser Pro Val His Pro
275 280 285
Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys Arg Pro Arg Gly
290 295 300
Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp
305 310 315 320
Ala Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu Ser Leu Ile Ile
325 330 335
Thr Leu Ile Cys Tyr His Arg Ser Arg Lys Arg Val Cys Lys Cys Pro
340 345 350
Arg Pro Leu Val Arg Gln Glu Gly Lys Pro Arg Pro Ser Glu Lys Ile
355 360 365
Val Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
370 375
<210> 39
<211> 1031
<212> DNA
<213> 嵌合抗原受体蛋白EpHL-C8-47编码核酸序列
<400> 39
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgct 60
cgtgatgacc cagactccac tctccctgcc tgtcagtctt ggagatcaag cctccatctc 120
ttgcagatct agtcagagcc ttgtacacag taatggaaac acctatttac attggtacct 180
gcagaagcca ggccagtctc caaagctcct gatctacaaa gtttccaacc gattttctgg 240
ggtcccagac aggttcagtg gcagtggatc agggacagat ttcacactca agatcagcag 300
agtggaggct gaggatctgg gagtttattt ctgctctcaa agtacacatg ttccgtacac 360
gttcggaggg gggaccaagc ttgagatcaa aggtggtggt ggttctggcg gcggcggctc 420
cggtggtggt ggttctgatc cttgagtgga ttggagaggt ttatcctaga attggtaatg 480
cttactacaa tgagaagttc aagggcaagg ccacactgac tgcagacaaa tcctccagca 540
cagcgtccat ggagctccgc agcctgacct ctgaggactc tgcggtctat ttctgtgcaa 600
gacggggatc ctacgatact aactacgact ggtacttcga tgtctggggc caagggacca 660
cggtcaccgt ctcctcaagt tctgtcgtgc cagtccttca gaaagtgaac tctactacta 720
ccaagccagt gctgcgaact ccctcacctg tgcaccctac cgggacatct cagccccaga 780
gaccagaaga ttgtcggccc cgtggctcag tgaaggggac cggattggac ttcgcctgtg 840
atatttacat ctgggcaccc ttggccggaa tctgcgtggc ccttctgctg tccttgatca 900
tcactctcat ctgctaccac aggagccgaa agcgtgtttg caaatgtccc aggccgctag 960
tcagacagga aggcaagccc agaccttcag agaaaattgt gtacccatac gatgttccag 1020
attacgctta a 1031
<210> 40
<211> 387
<212> PRT
<213> 嵌合抗原受体蛋白EpHL-C8-47氨基酸序列
<400> 40
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Leu Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val
20 25 30
Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu
35 40 45
Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro
50 55 60
Gly Gln Ser Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser
65 70 75 80
Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
85 90 95
Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys
100 105 110
Ser Gln Ser Thr His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu
115 120 125
Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Ala Arg
145 150 155 160
Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe
165 170 175
Thr Asn Tyr Gly Leu Ser Trp Val Lys Gln Arg Pro Gly Gln Val Leu
180 185 190
Glu Trp Ile Gly Glu Val Tyr Pro Arg Ile Gly Asn Ala Tyr Tyr Asn
195 200 205
Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser
210 215 220
Thr Ala Ser Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val
225 230 235 240
Tyr Phe Cys Ala Arg Arg Gly Ser Tyr Asp Thr Asn Tyr Asp Trp Tyr
245 250 255
Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ser Ser
260 265 270
Val Val Pro Val Leu Gln Lys Val Asn Ser Thr Thr Thr Lys Pro Val
275 280 285
Leu Arg Thr Pro Ser Pro Val His Pro Thr Gly Thr Ser Gln Pro Gln
290 295 300
Arg Pro Glu Asp Cys Arg Pro Arg Gly Ser Val Lys Gly Thr Gly Leu
305 310 315 320
Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Ile Cys
325 330 335
Val Ala Leu Leu Leu Ser Leu Ile Ile Thr Leu Ile Cys Tyr His Arg
340 345 350
Ser Arg Lys Arg Val Cys Lys Cys Pro Arg Pro Leu Val Arg Gln Glu
355 360 365
Gly Lys Pro Arg Pro Ser Glu Lys Ile Val Tyr Pro Tyr Asp Val Pro
370 375 380
Asp Tyr Ala
385
<210> 41
<211> 1031
<212> DNA
<213> 嵌合抗原受体蛋白EpLH-C8-47编码核酸序列
<400> 41
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactcctcc 60
ttgagtggat tggagaggtt tatcctagaa ttggtaatgc ttactacaat gagaagttca 120
agggcaaggc cacactgact gcagacaaat cctccagcac agcgtccatg gagctccgca 180
gcctgacctc tgaggactct gcggtctatt tctgtgcaag acggggatcc tacgatacta 240
actacgactg gtacttcgat gtctggggcc aagggaccac ggtcaccgtc tcctcaggtg 300
gtggtggttc tggcggcggc ggctccggtg gtggtggttc tgagctcgtg atgacccaga 360
ctccactctc cctgcctgtc agtcttggag atcaagcctc catctcttgc agatctagtc 420
agagccttgt acacagtaat ggaaacacct atttacattg gtacctgcag aagccaggcc 480
agtctccaaa gctcctgatc tacaaagttt ccaaccgatt ttctggggtc ccagacaggt 540
tcagtggcag tggatcaggg acagatttca cactcaagat cagcagagtg gaggctgagg 600
atctgggagt ttatttctgc tctcaaagta cacatgttcc gtacacgttc ggagggggga 660
ccaagcttga gatcaaaagt tctgtcgtgc cagtccttca gaaagtgaac tctactacta 720
ccaagccagt gctgcgaact ccctcacctg tgcaccctac cgggacatct cagccccaga 780
gaccagaaga ttgtcggccc cgtggctcag tgaaggggac cggattggac ttcgcctgtg 840
atatttacat ctgggcaccc ttggccggaa tctgcgtggc ccttctgctg tccttgatca 900
tcactctcat ctgctaccac aggagccgaa agcgtgtttg caaatgtccc aggccgctag 960
tcagacagga aggcaagccc agaccttcag agaaaattgt gtacccatac gatgttccag 1020
attacgctta a 1031
<210> 42
<211> 387
<212> PRT
<213> 嵌合抗原受体蛋白EpLH-C8-47氨基酸序列
<400> 42
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Ala
20 25 30
Arg Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr
35 40 45
Phe Thr Asn Tyr Gly Leu Ser Trp Val Lys Gln Arg Pro Gly Gln Val
50 55 60
Leu Glu Trp Ile Gly Glu Val Tyr Pro Arg Ile Gly Asn Ala Tyr Tyr
65 70 75 80
Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Ser Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala
100 105 110
Val Tyr Phe Cys Ala Arg Arg Gly Ser Tyr Asp Thr Asn Tyr Asp Trp
115 120 125
Tyr Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly
130 135 140
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu
145 150 155 160
Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly Asp Gln
165 170 175
Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly
180 185 190
Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Lys
195 200 205
Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp Arg
210 215 220
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg
225 230 235 240
Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser Thr His
245 250 255
Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Ser Ser
260 265 270
Val Val Pro Val Leu Gln Lys Val Asn Ser Thr Thr Thr Lys Pro Val
275 280 285
Leu Arg Thr Pro Ser Pro Val His Pro Thr Gly Thr Ser Gln Pro Gln
290 295 300
Arg Pro Glu Asp Cys Arg Pro Arg Gly Ser Val Lys Gly Thr Gly Leu
305 310 315 320
Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Ile Cys
325 330 335
Val Ala Leu Leu Leu Ser Leu Ile Ile Thr Leu Ile Cys Tyr His Arg
340 345 350
Ser Arg Lys Arg Val Cys Lys Cys Pro Arg Pro Leu Val Arg Gln Glu
355 360 365
Gly Lys Pro Arg Pro Ser Glu Lys Ile Val Tyr Pro Tyr Asp Val Pro
370 375 380
Asp Tyr Ala
385
<210> 43
<211> 1149
<212> DNA
<213> 嵌合抗原受体蛋白EpHL-C8-510编码核酸序列
<400> 43
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgag 60
gtgcagctgc tcgagcagtc tggagctgag ctggtaaggc ctgggacttc agtgaagata 120
tcctgcaagg cttctggata cgccttcact aactactggc taggttgggt aaagcagagg 180
cctggacatg gacttgagtg gattggagat attttccctg gaagtggtaa tatccactac 240
aatgagaagt tcaagggcaa agccacactg actgcagaca aatcttcgag cacagcctat 300
atgcagctca gtagcctgac atttgaggac tctgctgtct atttctgtgc aagactgagg 360
aactgggacg agcctatgga ctactggggc caagggacca cggtcaccgt cggtggtggt 420
ggttctggcg gcggcggctc cggtggtggt ggttctgagc tcgtgatgac acagtctcca 480
tcctccctga ctgtgacagc aggagagaag gtcactatga gctgcaagtc cagtcagagt 540
ctgttaaaca gtggaaatca aaagaactac ttgacctggt accagcagaa accagggcag 600
cctcctaaac tgttgatcta ctgggcatcc actagggaat ctggggtccc tgatcgcttc 660
acaggcagtg gatctggaac agatttcact ctcaccatca gcagtgtgca ggctgaagac 720
ctggcagttt attactgtca gaatgattat agttatccgc tcacgttcgg tgctgggacc 780
aagcttgaga tcaaaagttc tgtcgtgcca gtccttcaga aagtgaactc tactactacc 840
aagccagtgc tgcgaactcc ctcacctgtg caccctaccg ggacatctca gccccagaga 900
ccagaagatt gtcggccccg tggctcagtg aaggggaccg gattggactt cgcctgtgat 960
atttacatct gggcaccctt ggccggaatc tgcgtggccc ttctgctgtc cttgatcatc 1020
actctcatct gctaccacag gagccgaaag cgtgtttgca aatgtcccag gccgctagtc 1080
agacaggaag gcaagcccag accttcagag aaaattgtgt acccatacga tgttccagat 1140
tacgcttaa 1149
<210> 44
<211> 382
<212> PRT
<213> 嵌合抗原受体蛋白EpHL-C8-510氨基酸序列
<400> 44
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val
20 25 30
Arg Pro Gly Thr Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala
35 40 45
Phe Thr Asn Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly
50 55 60
Leu Glu Trp Ile Gly Asp Ile Phe Pro Gly Ser Gly Asn Ile His Tyr
65 70 75 80
Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Phe Glu Asp Ser Ala
100 105 110
Val Tyr Phe Cys Ala Arg Leu Arg Asn Trp Asp Glu Pro Met Asp Tyr
115 120 125
Trp Gly Gln Gly Thr Thr Val Thr Val Gly Gly Gly Gly Ser Gly Gly
130 135 140
Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Val Met Thr Gln Ser Pro
145 150 155 160
Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys
165 170 175
Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu Thr
180 185 190
Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp
195 200 205
Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly
210 215 220
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp
225 230 235 240
Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Ser Tyr Pro Leu Thr Phe
245 250 255
Gly Ala Gly Thr Lys Leu Glu Ile Lys Ser Ser Val Val Pro Val Leu
260 265 270
Gln Lys Val Asn Ser Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser
275 280 285
Pro Val His Pro Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys
290 295 300
Arg Pro Arg Gly Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp
305 310 315 320
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu
325 330 335
Ser Leu Ile Ile Thr Leu Ile Cys Tyr His Arg Ser Arg Lys Arg Val
340 345 350
Cys Lys Cys Pro Arg Pro Leu Val Arg Gln Glu Gly Lys Pro Arg Pro
355 360 365
Ser Glu Lys Ile Val Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
370 375 380
<210> 45
<211> 1149
<212> DNA
<213> 嵌合抗原受体蛋白EpLH-C8-510编码核酸序列
<400> 45
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgag 60
ctcgtgatga cacagtctcc atcctccctg actgtgacag caggagagaa ggtcactatg 120
agctgcaagt ccagtcagag tctgttaaac agtggaaatc aaaagaacta cttgacctgg 180
taccagcaga aaccagggca gcctcctaaa ctgttgatct actgggcatc cactagggaa 240
tctggggtcc ctgatcgctt cacaggcagt ggatctggaa cagatttcac tctcaccatc 300
agcagtgtgc aggctgaaga cctggcagtt tattactgtc agaatgatta tagttatccg 360
ctcacgttcg gtgctgggac caagcttgag atcaaaggtg gtggtggttc tggcggcggc 420
ggctccggtg gtggtggttc tgaggtgcag ctgctcgagc agtctggagc tgagctggta 480
aggcctggga cttcagtgaa gatatcctgc aaggcttctg gatacgcctt cactaactac 540
tggctaggtt gggtaaagca gaggcctgga catggacttg agtggattgg agatattttc 600
cctggaagtg gtaatatcca ctacaatgag aagttcaagg gcaaagccac actgactgca 660
gacaaatctt cgagcacagc ctatatgcag ctcagtagcc tgacatttga ggactctgct 720
gtctatttct gtgcaagact gaggaactgg gacgagccta tggactactg gggccaaggg 780
accacggtca ccgtcagttc tgtcgtgcca gtccttcaga aagtgaactc tactactacc 840
aagccagtgc tgcgaactcc ctcacctgtg caccctaccg ggacatctca gccccagaga 900
ccagaagatt gtcggccccg tggctcagtg aaggggaccg gattggactt cgcctgtgat 960
atttacatct gggcaccctt ggccggaatc tgcgtggccc ttctgctgtc cttgatcatc 1020
actctcatct gctaccacag gagccgaaag cgtgtttgca aatgtcccag gccgctagtc 1080
agacaggaag gcaagcccag accttcagag aaaattgtgt acccatacga tgttccagat 1140
tacgcttaa 1149
<210> 46
<211> 382
<212> PRT
<213> 嵌合抗原受体蛋白EpLH-C8-510氨基酸序列
<400> 46
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Leu Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val
20 25 30
Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu
35 40 45
Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys
50 55 60
Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe
85 90 95
Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr
100 105 110
Cys Gln Asn Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys
115 120 125
Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
130 135 140
Gly Gly Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val
145 150 155 160
Arg Pro Gly Thr Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala
165 170 175
Phe Thr Asn Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly
180 185 190
Leu Glu Trp Ile Gly Asp Ile Phe Pro Gly Ser Gly Asn Ile His Tyr
195 200 205
Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
210 215 220
Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Phe Glu Asp Ser Ala
225 230 235 240
Val Tyr Phe Cys Ala Arg Leu Arg Asn Trp Asp Glu Pro Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Val Val Pro Val Leu
260 265 270
Gln Lys Val Asn Ser Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser
275 280 285
Pro Val His Pro Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys
290 295 300
Arg Pro Arg Gly Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp
305 310 315 320
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu
325 330 335
Ser Leu Ile Ile Thr Leu Ile Cys Tyr His Arg Ser Arg Lys Arg Val
340 345 350
Cys Lys Cys Pro Arg Pro Leu Val Arg Gln Glu Gly Lys Pro Arg Pro
355 360 365
Ser Glu Lys Ile Val Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
370 375 380
<210> 47
<211> 1530
<212> DNA
<213> 嵌合抗原受体蛋白EpHL-GP-31编码核酸序列
<400> 47
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgct 60
cgtcatgacc cagtctccat cttatcttgc tgcatctcct ggagaaacca ttactattaa 120
ttgcagggca agtaagagca ttagcaaata tttagcctgg tatcaagaga aacctgggaa 180
aactaataag cttcttatct actctggatc cactttgcaa tctggaattc catcaaggtt 240
cagtggcagt ggatctggta cagatttcac tctcaccatc agtagcctgg agcctgaaga 300
ttttgcaatg tattactgtc aacagcataa tgaatatccg tacacgttcg gaggggggac 360
caagcttgag atcaaaggtg gtggtggttc tggcggcggc ggctccggtg gtggtggttc 420
tgagaggtgc agctgctcga gcagtctgga gctgagctgg tgaaacctgg ggcctcagtg 480
aagatatcct gcaaggcttc tggatacgcc ttcactaact actggctagg ttgggtaaag 540
cagaggcctg gacatggact tgagtggatt ggagatcttt tccctggaag tggtaatact 600
cactacaatg agaggttcag gggcaaagcc acactgactg cagacaaatc ctcgagcaca 660
gcctttatgc agctcagtag cctgacatct gaggactctg ctgtctattt ctgtgcaaga 720
ttgaggaact gggacgaggc tatggactac tggggccaag ggaccacggt caccgtctcc 780
tcaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 840
gacggcgacg taaacggcca caagttcagc gtgtctggcg agggcgaggg cgatgccacc 900
tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 960
accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 1020
aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 1080
ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 1140
ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 1200
cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 1260
aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 1320
gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 1380
cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 1440
gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 1500
tacccatacg atgttccaga ttacgcttaa 1530
<210> 48
<211> 509
<212> PRT
<213> 嵌合抗原受体蛋白EpHL-GP-31氨基酸序列
<400> 48
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Leu Val Met Thr Gln Ser Pro Ser Tyr Leu Ala Ala
20 25 30
Ser Pro Gly Glu Thr Ile Thr Ile Asn Cys Arg Ala Ser Lys Ser Ile
35 40 45
Ser Lys Tyr Leu Ala Trp Tyr Gln Glu Lys Pro Gly Lys Thr Asn Lys
50 55 60
Leu Leu Ile Tyr Ser Gly Ser Thr Leu Gln Ser Gly Ile Pro Ser Arg
65 70 75 80
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
85 90 95
Leu Glu Pro Glu Asp Phe Ala Met Tyr Tyr Cys Gln Gln His Asn Glu
100 105 110
Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln
130 135 140
Leu Leu Glu Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val
145 150 155 160
Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Trp Leu
165 170 175
Gly Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile Gly Asp
180 185 190
Leu Phe Pro Gly Ser Gly Asn Thr His Tyr Asn Glu Arg Phe Arg Gly
195 200 205
Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Phe Met Gln
210 215 220
Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg
225 230 235 240
Leu Arg Asn Trp Asp Glu Ala Met Asp Tyr Trp Gly Gln Gly Thr Thr
245 250 255
Val Thr Val Ser Ser Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly
260 265 270
Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn Gly His Lys
275 280 285
Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu
290 295 300
Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro
305 310 315 320
Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr
325 330 335
Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu
340 345 350
Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr
355 360 365
Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg
370 375 380
Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly
385 390 395 400
His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala
405 410 415
Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn
420 425 430
Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr
435 440 445
Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser
450 455 460
Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met
465 470 475 480
Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp
485 490 495
Glu Leu Tyr Lys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
500 505
<210> 49
<211> 1530
<212> DNA
<213> 嵌合抗原受体蛋白EpLH-GP-31编码核酸序列
<400> 49
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgag 60
gtgcagctgc tcgagcagtc tggagctgag ctggtgaaac ctggggcctc agtgaagata 120
tcctgcaagg cttctggata cgccttcact aactactggc taggttgggt aaagcagagg 180
cctggacatg gacttgagtg gattggagat cttttccctg gaagtggtaa tactcactac 240
aatgagaggt tcaggggcaa agccacactg actgcagaca aatcctcgag cacagccttt 300
atgcagctca gtagcctgac atctgaggac tctgctgtct atttctgtgc aagattgagg 360
aactgggacg aggctatgga ctactggggc caagggacca cggtcaccgt ctcctcaggt 420
ggtggtggtt ctggcggcgg cggctccggt ggtggtggtt ctgagctcgt catgacccag 480
tctccatctt atcttgctgc atctcctgga gaaaccatta ctattaattg cagggcaagt 540
aagagcatta gcaaatattt agcctggtat caagagaaac ctgggaaaac taataagctt 600
cttatctact ctggatccac tttgcaatct ggaattccat caaggttcag tggcagtgga 660
tctggtacag atttcactct caccatcagt agcctggagc ctgaagattt tgcaatgtat 720
tactgtcaac agcataatga atatccgtac acgttcggag gggggaccaa gcttgagatc 780
aaaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 840
gacggcgacg taaacggcca caagttcagc gtgtctggcg agggcgaggg cgatgccacc 900
tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 960
accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 1020
aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 1080
ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 1140
ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 1200
cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 1260
aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 1320
gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 1380
cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 1440
gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 1500
tacccatacg atgttccaga ttacgcttaa 1530
<210> 50
<211> 509
<212> PRT
<213> 嵌合抗原受体蛋白EpLH-GP-31氨基酸序列
<400> 50
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val
20 25 30
Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala
35 40 45
Phe Thr Asn Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly
50 55 60
Leu Glu Trp Ile Gly Asp Leu Phe Pro Gly Ser Gly Asn Thr His Tyr
65 70 75 80
Asn Glu Arg Phe Arg Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Phe Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala
100 105 110
Val Tyr Phe Cys Ala Arg Leu Arg Asn Trp Asp Glu Ala Met Asp Tyr
115 120 125
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser
130 135 140
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Val Met Thr Gln
145 150 155 160
Ser Pro Ser Tyr Leu Ala Ala Ser Pro Gly Glu Thr Ile Thr Ile Asn
165 170 175
Cys Arg Ala Ser Lys Ser Ile Ser Lys Tyr Leu Ala Trp Tyr Gln Glu
180 185 190
Lys Pro Gly Lys Thr Asn Lys Leu Leu Ile Tyr Ser Gly Ser Thr Leu
195 200 205
Gln Ser Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
210 215 220
Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Met Tyr
225 230 235 240
Tyr Cys Gln Gln His Asn Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr
245 250 255
Lys Leu Glu Ile Lys Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly
260 265 270
Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn Gly His Lys
275 280 285
Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu
290 295 300
Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro
305 310 315 320
Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr
325 330 335
Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu
340 345 350
Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr
355 360 365
Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg
370 375 380
Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly
385 390 395 400
His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala
405 410 415
Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn
420 425 430
Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr
435 440 445
Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser
450 455 460
Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met
465 470 475 480
Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp
485 490 495
Glu Leu Tyr Lys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
500 505
<210> 51
<211> 1424
<212> DNA
<213> 嵌合抗原受体蛋白EpHL-GP-47编码核酸序列
<400> 51
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgct 60
cgtgatgacc cagactccac tctccctgcc tgtcagtctt ggagatcaag cctccatctc 120
ttgcagatct agtcagagcc ttgtacacag taatggaaac acctatttac attggtacct 180
gcagaagcca ggccagtctc caaagctcct gatctacaaa gtttccaacc gattttctgg 240
ggtcccagac aggttcagtg gcagtggatc agggacagat ttcacactca agatcagcag 300
agtggaggct gaggatctgg gagtttattt ctgctctcaa agtacacatg ttccgtacac 360
gttcggaggg gggaccaagc ttgagatcaa aggtggtggt ggttctggcg gcggcggctc 420
cggtggtggt ggttctgatc cttgagtgga ttggagaggt ttatcctaga attggtaatg 480
cttactacaa tgagaagttc aagggcaagg ccacactgac tgcagacaaa tcctccagca 540
cagcgtccat ggagctccgc agcctgacct ctgaggactc tgcggtctat ttctgtgcaa 600
gacggggatc ctacgatact aactacgact ggtacttcga tgtctggggc caagggacca 660
cggtcaccgt ctcctcaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca 720
tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg 780
agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc 840
ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct 900
accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc 960
aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt 1020
tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg 1080
gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg 1140
ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg 1200
gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 1260
tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga 1320
agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg 1380
acgagctgta caagtaccca tacgatgttc cagattacgc ttaa 1424
<210> 52
<211> 518
<212> PRT
<213> 嵌合抗原受体蛋白EpHL-GP-47氨基酸序列
<400> 52
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Leu Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val
20 25 30
Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu
35 40 45
Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro
50 55 60
Gly Gln Ser Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser
65 70 75 80
Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
85 90 95
Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys
100 105 110
Ser Gln Ser Thr His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu
115 120 125
Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
130 135 140
Gly Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Ala Arg
145 150 155 160
Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe
165 170 175
Thr Asn Tyr Gly Leu Ser Trp Val Lys Gln Arg Pro Gly Gln Val Leu
180 185 190
Glu Trp Ile Gly Glu Val Tyr Pro Arg Ile Gly Asn Ala Tyr Tyr Asn
195 200 205
Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser
210 215 220
Thr Ala Ser Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val
225 230 235 240
Tyr Phe Cys Ala Arg Arg Gly Ser Tyr Asp Thr Asn Tyr Asp Trp Tyr
245 250 255
Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Met Val
260 265 270
Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu
275 280 285
Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly
290 295 300
Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr
305 310 315 320
Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu Thr
325 330 335
Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His
340 345 350
Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr
355 360 365
Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys
370 375 380
Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp
385 390 395 400
Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr
405 410 415
Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile
420 425 430
Lys Ala Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln
435 440 445
Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val
450 455 460
Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys
465 470 475 480
Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr
485 490 495
Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys Tyr Pro Tyr
500 505 510
Asp Val Pro Asp Tyr Ala
515
<210> 53
<211> 1424
<212> DNA
<213> 嵌合抗原受体蛋白EpLH-GP-47编码核酸序列
<400> 53
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactcctcc 60
ttgagtggat tggagaggtt tatcctagaa ttggtaatgc ttactacaat gagaagttca 120
agggcaaggc cacactgact gcagacaaat cctccagcac agcgtccatg gagctccgca 180
gcctgacctc tgaggactct gcggtctatt tctgtgcaag acggggatcc tacgatacta 240
actacgactg gtacttcgat gtctggggcc aagggaccac ggtcaccgtc tcctcaggtg 300
gtggtggttc tggcggcggc ggctccggtg gtggtggttc tgagctcgtg atgacccaga 360
ctccactctc cctgcctgtc agtcttggag atcaagcctc catctcttgc agatctagtc 420
agagccttgt acacagtaat ggaaacacct atttacattg gtacctgcag aagccaggcc 480
agtctccaaa gctcctgatc tacaaagttt ccaaccgatt ttctggggtc ccagacaggt 540
tcagtggcag tggatcaggg acagatttca cactcaagat cagcagagtg gaggctgagg 600
atctgggagt ttatttctgc tctcaaagta cacatgttcc gtacacgttc ggagggggga 660
ccaagcttga gatcaaaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca 720
tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg 780
agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc 840
ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct 900
accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc 960
aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt 1020
tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg 1080
gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg 1140
ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg 1200
gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 1260
tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga 1320
agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg 1380
acgagctgta caagtaccca tacgatgttc cagattacgc ttaa 1424
<210> 54
<211> 518
<212> PRT
<213> 嵌合抗原受体蛋白EpLH-GP-47氨基酸序列
<400> 54
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Ala
20 25 30
Arg Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr
35 40 45
Phe Thr Asn Tyr Gly Leu Ser Trp Val Lys Gln Arg Pro Gly Gln Val
50 55 60
Leu Glu Trp Ile Gly Glu Val Tyr Pro Arg Ile Gly Asn Ala Tyr Tyr
65 70 75 80
Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Ser Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala
100 105 110
Val Tyr Phe Cys Ala Arg Arg Gly Ser Tyr Asp Thr Asn Tyr Asp Trp
115 120 125
Tyr Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly
130 135 140
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu
145 150 155 160
Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly Asp Gln
165 170 175
Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly
180 185 190
Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Lys
195 200 205
Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp Arg
210 215 220
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg
225 230 235 240
Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser Thr His
245 250 255
Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Met Val
260 265 270
Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu
275 280 285
Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly
290 295 300
Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr
305 310 315 320
Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu Thr
325 330 335
Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His
340 345 350
Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr
355 360 365
Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys
370 375 380
Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp
385 390 395 400
Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr
405 410 415
Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile
420 425 430
Lys Ala Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln
435 440 445
Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val
450 455 460
Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys
465 470 475 480
Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr
485 490 495
Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys Tyr Pro Tyr
500 505 510
Asp Val Pro Asp Tyr Ala
515
<210> 55
<211> 1542
<212> DNA
<213> 嵌合抗原受体蛋白EpHL-GP-510编码核酸序列
<400> 55
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgag 60
gtgcagctgc tcgagcagtc tggagctgag ctggtaaggc ctgggacttc agtgaagata 120
tcctgcaagg cttctggata cgccttcact aactactggc taggttgggt aaagcagagg 180
cctggacatg gacttgagtg gattggagat attttccctg gaagtggtaa tatccactac 240
aatgagaagt tcaagggcaa agccacactg actgcagaca aatcttcgag cacagcctat 300
atgcagctca gtagcctgac atttgaggac tctgctgtct atttctgtgc aagactgagg 360
aactgggacg agcctatgga ctactggggc caagggacca cggtcaccgt cggtggtggt 420
ggttctggcg gcggcggctc cggtggtggt ggttctgagc tcgtgatgac acagtctcca 480
tcctccctga ctgtgacagc aggagagaag gtcactatga gctgcaagtc cagtcagagt 540
ctgttaaaca gtggaaatca aaagaactac ttgacctggt accagcagaa accagggcag 600
cctcctaaac tgttgatcta ctgggcatcc actagggaat ctggggtccc tgatcgcttc 660
acaggcagtg gatctggaac agatttcact ctcaccatca gcagtgtgca ggctgaagac 720
ctggcagttt attactgtca gaatgattat agttatccgc tcacgttcgg tgctgggacc 780
aagcttgaga tcaaaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc 840
ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag 900
ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc 960
gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac 1020
cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag 1080
gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc 1140
gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc 1200
aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc 1260
gacaagcaga agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc 1320
agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg 1380
ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag 1440
cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac 1500
gagctgtaca agtacccata cgatgttcca gattacgctt aa 1542
<210> 56
<211> 513
<212> PRT
<213> 嵌合抗原受体蛋白EpHL-GP-510氨基酸序列
<400> 56
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val
20 25 30
Arg Pro Gly Thr Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala
35 40 45
Phe Thr Asn Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly
50 55 60
Leu Glu Trp Ile Gly Asp Ile Phe Pro Gly Ser Gly Asn Ile His Tyr
65 70 75 80
Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Phe Glu Asp Ser Ala
100 105 110
Val Tyr Phe Cys Ala Arg Leu Arg Asn Trp Asp Glu Pro Met Asp Tyr
115 120 125
Trp Gly Gln Gly Thr Thr Val Thr Val Gly Gly Gly Gly Ser Gly Gly
130 135 140
Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Val Met Thr Gln Ser Pro
145 150 155 160
Ser Ser Leu Thr Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys
165 170 175
Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu Thr
180 185 190
Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp
195 200 205
Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly
210 215 220
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp
225 230 235 240
Leu Ala Val Tyr Tyr Cys Gln Asn Asp Tyr Ser Tyr Pro Leu Thr Phe
245 250 255
Gly Ala Gly Thr Lys Leu Glu Ile Lys Met Val Ser Lys Gly Glu Glu
260 265 270
Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val
275 280 285
Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr
290 295 300
Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro
305 310 315 320
Val Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys
325 330 335
Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser
340 345 350
Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp
355 360 365
Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr
370 375 380
Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly
385 390 395 400
Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn Val
405 410 415
Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys
420 425 430
Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr
435 440 445
Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn
450 455 460
His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys
465 470 475 480
Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr
485 490 495
Leu Gly Met Asp Glu Leu Tyr Lys Tyr Pro Tyr Asp Val Pro Asp Tyr
500 505 510
Ala
<210> 57
<211> 1542
<212> DNA
<213> 嵌合抗原受体蛋白EpLH-GP-510编码核酸序列
<400> 57
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt acactccgag 60
ctcgtgatga cacagtctcc atcctccctg actgtgacag caggagagaa ggtcactatg 120
agctgcaagt ccagtcagag tctgttaaac agtggaaatc aaaagaacta cttgacctgg 180
taccagcaga aaccagggca gcctcctaaa ctgttgatct actgggcatc cactagggaa 240
tctggggtcc ctgatcgctt cacaggcagt ggatctggaa cagatttcac tctcaccatc 300
agcagtgtgc aggctgaaga cctggcagtt tattactgtc agaatgatta tagttatccg 360
ctcacgttcg gtgctgggac caagcttgag atcaaaggtg gtggtggttc tggcggcggc 420
ggctccggtg gtggtggttc tgaggtgcag ctgctcgagc agtctggagc tgagctggta 480
aggcctggga cttcagtgaa gatatcctgc aaggcttctg gatacgcctt cactaactac 540
tggctaggtt gggtaaagca gaggcctgga catggacttg agtggattgg agatattttc 600
cctggaagtg gtaatatcca ctacaatgag aagttcaagg gcaaagccac actgactgca 660
gacaaatctt cgagcacagc ctatatgcag ctcagtagcc tgacatttga ggactctgct 720
gtctatttct gtgcaagact gaggaactgg gacgagccta tggactactg gggccaaggg 780
accacggtca ccgtcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc 840
ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag 900
ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc 960
gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac 1020
cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag 1080
gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc 1140
gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc 1200
aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc 1260
gacaagcaga agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc 1320
agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg 1380
ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag 1440
cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac 1500
gagctgtaca agtacccata cgatgttcca gattacgctt aa 1542
<210> 58
<211> 513
<212> PRT
<213> 嵌合抗原受体蛋白EpLH-GP-510氨基酸序列
<400> 58
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly
1 5 10 15
Val His Ser Glu Leu Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val
20 25 30
Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu
35 40 45
Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys
50 55 60
Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe
85 90 95
Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr
100 105 110
Cys Gln Asn Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys
115 120 125
Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
130 135 140
Gly Gly Ser Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Leu Val
145 150 155 160
Arg Pro Gly Thr Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala
165 170 175
Phe Thr Asn Tyr Trp Leu Gly Trp Val Lys Gln Arg Pro Gly His Gly
180 185 190
Leu Glu Trp Ile Gly Asp Ile Phe Pro Gly Ser Gly Asn Ile His Tyr
195 200 205
Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
210 215 220
Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Phe Glu Asp Ser Ala
225 230 235 240
Val Tyr Phe Cys Ala Arg Leu Arg Asn Trp Asp Glu Pro Met Asp Tyr
245 250 255
Trp Gly Gln Gly Thr Thr Val Thr Val Met Val Ser Lys Gly Glu Glu
260 265 270
Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val
275 280 285
Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr
290 295 300
Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro
305 310 315 320
Val Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys
325 330 335
Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser
340 345 350
Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp
355 360 365
Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr
370 375 380
Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly
385 390 395 400
Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn Val
405 410 415
Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys
420 425 430
Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr
435 440 445
Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn
450 455 460
His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys
465 470 475 480
Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr
485 490 495
Leu Gly Met Asp Glu Leu Tyr Lys Tyr Pro Tyr Asp Val Pro Asp Tyr
500 505 510
Ala
<210> 59
<211> 21
<212> DNA
<213> hCDH1-F荧光定量PCR引物
<400> 59
gaacagcacg tacacagccc t 21
<210> 60
<211> 21
<212> DNA
<213> hCDH1-R荧光定量PCR引物
<400> 60
gcagaagtgt ccctgttcca g 21
<210> 61
<211> 19
<212> DNA
<213> hGAPDH-F荧光定量PCR引物
<400> 61
gaaggtgaag gtcggagtc 19
<210> 62
<211> 20
<212> DNA
<213> hGAPDH-R荧光定量PCR引物
<400> 62
gaagatggtg atgggatttc 20