CN113122617B - 一种筛选特异bcr/tcr的方法及其系统 - Google Patents
一种筛选特异bcr/tcr的方法及其系统 Download PDFInfo
- Publication number
- CN113122617B CN113122617B CN202110273631.0A CN202110273631A CN113122617B CN 113122617 B CN113122617 B CN 113122617B CN 202110273631 A CN202110273631 A CN 202110273631A CN 113122617 B CN113122617 B CN 113122617B
- Authority
- CN
- China
- Prior art keywords
- bcr
- tcr
- sequences
- sample
- peripheral blood
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000012216 screening Methods 0.000 title claims abstract description 36
- 238000000034 method Methods 0.000 title claims abstract description 31
- 108091007433 antigens Proteins 0.000 claims abstract description 26
- 102000036639 antigens Human genes 0.000 claims abstract description 26
- 239000000427 antigen Substances 0.000 claims abstract description 25
- 238000012163 sequencing technique Methods 0.000 claims abstract description 24
- 210000004698 lymphocyte Anatomy 0.000 claims abstract description 15
- 238000010839 reverse transcription Methods 0.000 claims abstract description 10
- 238000012165 high-throughput sequencing Methods 0.000 claims abstract description 9
- 108020004999 messenger RNA Proteins 0.000 claims abstract description 9
- 230000003321 amplification Effects 0.000 claims abstract description 8
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 8
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 8
- 238000007403 mPCR Methods 0.000 claims abstract description 7
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 6
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 6
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 6
- 230000000638 stimulation Effects 0.000 claims abstract description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 210000005259 peripheral blood Anatomy 0.000 claims description 88
- 239000011886 peripheral blood Substances 0.000 claims description 88
- 239000000523 sample Substances 0.000 claims description 34
- 101100112922 Candida albicans CDR3 gene Proteins 0.000 claims description 23
- 230000002516 postimmunization Effects 0.000 claims description 6
- 206010028980 Neoplasm Diseases 0.000 claims description 3
- 238000000338 in vitro Methods 0.000 claims description 3
- 210000000952 spleen Anatomy 0.000 claims description 3
- 210000001519 tissue Anatomy 0.000 claims description 3
- 239000012805 animal sample Substances 0.000 claims 1
- 108091008874 T cell receptors Proteins 0.000 abstract description 33
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 abstract description 31
- 230000003053 immunization Effects 0.000 abstract description 8
- 238000002649 immunization Methods 0.000 abstract description 8
- 108091008875 B cell receptors Proteins 0.000 description 32
- 241000711573 Coronaviridae Species 0.000 description 19
- 210000004369 blood Anatomy 0.000 description 17
- 239000008280 blood Substances 0.000 description 17
- 238000003752 polymerase chain reaction Methods 0.000 description 16
- 210000004027 cell Anatomy 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 12
- 108010068265 aspartyltyrosine Proteins 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 6
- 150000001413 amino acids Chemical group 0.000 description 6
- 150000002500 ions Chemical class 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 210000003719 b-lymphocyte Anatomy 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 5
- 239000013610 patient sample Substances 0.000 description 5
- SDZRIBWEVVRDQI-CIUDSAMLSA-N Ala-Lys-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O SDZRIBWEVVRDQI-CIUDSAMLSA-N 0.000 description 4
- FVKRQMQQFGBXHV-QXEWZRGKSA-N Met-Asp-Val Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O FVKRQMQQFGBXHV-QXEWZRGKSA-N 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- LMKKMCGTDANZTR-BZSNNMDCSA-N Tyr-Phe-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=C(O)C=C1 LMKKMCGTDANZTR-BZSNNMDCSA-N 0.000 description 4
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 108010003137 tyrosyltyrosine Proteins 0.000 description 4
- SVBXIUDNTRTKHE-CIUDSAMLSA-N Ala-Arg-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O SVBXIUDNTRTKHE-CIUDSAMLSA-N 0.000 description 3
- JBGSZRYCXBPWGX-BQBZGAKWSA-N Ala-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CCCN=C(N)N JBGSZRYCXBPWGX-BQBZGAKWSA-N 0.000 description 3
- KOWYNSKRPUWSFG-IHPCNDPISA-N Asp-Phe-Trp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)O)NC(=O)[C@H](CC(=O)O)N KOWYNSKRPUWSFG-IHPCNDPISA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- OQPNSDWGAMFJNU-QWRGUYRKSA-N Ser-Gly-Tyr Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 OQPNSDWGAMFJNU-QWRGUYRKSA-N 0.000 description 3
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- DJDSEDOKJTZBAR-ZDLURKLDSA-N Thr-Gly-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O DJDSEDOKJTZBAR-ZDLURKLDSA-N 0.000 description 3
- 239000011324 bead Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000007405 data analysis Methods 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 108010087823 glycyltyrosine Proteins 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- QDRGPQWIVZNJQD-CIUDSAMLSA-N Ala-Arg-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O QDRGPQWIVZNJQD-CIUDSAMLSA-N 0.000 description 2
- KVWLTGNCJYDJET-LSJOCFKGSA-N Ala-Arg-His Chemical compound C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N KVWLTGNCJYDJET-LSJOCFKGSA-N 0.000 description 2
- MFMDKJIPHSWSBM-GUBZILKMSA-N Ala-Lys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O MFMDKJIPHSWSBM-GUBZILKMSA-N 0.000 description 2
- NZGRHTKZFSVPAN-BIIVOSGPSA-N Ala-Ser-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N NZGRHTKZFSVPAN-BIIVOSGPSA-N 0.000 description 2
- QLSRIZIDQXDQHK-RCWTZXSCSA-N Arg-Val-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QLSRIZIDQXDQHK-RCWTZXSCSA-N 0.000 description 2
- HPBNLFLSSQDFQW-WHFBIAKZSA-N Asn-Ser-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O HPBNLFLSSQDFQW-WHFBIAKZSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 208000001528 Coronaviridae Infections Diseases 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- FOBUGKUBUJOWAD-IHPCNDPISA-N Leu-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 FOBUGKUBUJOWAD-IHPCNDPISA-N 0.000 description 2
- YIRIDPUGZKHMHT-ACRUOGEOSA-N Leu-Tyr-Tyr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O YIRIDPUGZKHMHT-ACRUOGEOSA-N 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- IUVYJBMTHARMIP-PCBIJLKTSA-N Phe-Asp-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O IUVYJBMTHARMIP-PCBIJLKTSA-N 0.000 description 2
- MQVFHOPCKNTHGT-MELADBBJSA-N Phe-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O MQVFHOPCKNTHGT-MELADBBJSA-N 0.000 description 2
- NAXPHWZXEXNDIW-JTQLQIEISA-N Phe-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 NAXPHWZXEXNDIW-JTQLQIEISA-N 0.000 description 2
- XFFIGWGYMUFCCQ-ULQDDVLXSA-N Pro-His-Tyr Chemical compound C1=CC(O)=CC=C1C[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)[C@H]1[NH2+]CCC1)CC1=CN=CN1 XFFIGWGYMUFCCQ-ULQDDVLXSA-N 0.000 description 2
- JFWDJFULOLKQFY-QWRGUYRKSA-N Ser-Gly-Phe Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JFWDJFULOLKQFY-QWRGUYRKSA-N 0.000 description 2
- RRVFEDGUXSYWOW-BZSNNMDCSA-N Ser-Phe-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O RRVFEDGUXSYWOW-BZSNNMDCSA-N 0.000 description 2
- FHXGMDRKJHKLKW-QWRGUYRKSA-N Ser-Tyr-Gly Chemical compound OC[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=C(O)C=C1 FHXGMDRKJHKLKW-QWRGUYRKSA-N 0.000 description 2
- AZGZDDNKFFUDEH-QWRGUYRKSA-N Tyr-Gly-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AZGZDDNKFFUDEH-QWRGUYRKSA-N 0.000 description 2
- BSCBBPKDVOZICB-KKUMJFAQSA-N Tyr-Leu-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O BSCBBPKDVOZICB-KKUMJFAQSA-N 0.000 description 2
- OJCISMMNNUNNJA-BZSNNMDCSA-N Tyr-Tyr-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=C(O)C=C1 OJCISMMNNUNNJA-BZSNNMDCSA-N 0.000 description 2
- JQOMHZMWQHXALX-FHWLQOOXSA-N Tyr-Tyr-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JQOMHZMWQHXALX-FHWLQOOXSA-N 0.000 description 2
- WYOBRXPIZVKNMF-IRXDYDNUSA-N Tyr-Tyr-Gly Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)NCC(O)=O)C1=CC=C(O)C=C1 WYOBRXPIZVKNMF-IRXDYDNUSA-N 0.000 description 2
- BUPRFDPUIJNOLS-UFYCRDLUSA-N Tyr-Tyr-Met Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCSC)C(O)=O BUPRFDPUIJNOLS-UFYCRDLUSA-N 0.000 description 2
- RMRFSFXLFWWAJZ-HJOGWXRNSA-N Tyr-Tyr-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 RMRFSFXLFWWAJZ-HJOGWXRNSA-N 0.000 description 2
- 108010011559 alanylphenylalanine Proteins 0.000 description 2
- 108010093581 aspartyl-proline Proteins 0.000 description 2
- 238000010370 cell cloning Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 108010089804 glycyl-threonine Proteins 0.000 description 2
- 108010084389 glycyltryptophan Proteins 0.000 description 2
- 210000004754 hybrid cell Anatomy 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000002823 phage display Methods 0.000 description 2
- 108010070409 phenylalanyl-glycyl-glycine Proteins 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 108010061238 threonyl-glycine Proteins 0.000 description 2
- TTXMOJWKNRJWQJ-FXQIFTODSA-N Ala-Arg-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CCCN=C(N)N TTXMOJWKNRJWQJ-FXQIFTODSA-N 0.000 description 1
- JAMAWBXXKFGFGX-KZVJFYERSA-N Ala-Arg-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JAMAWBXXKFGFGX-KZVJFYERSA-N 0.000 description 1
- DWINFPQUSSHSFS-UVBJJODRSA-N Ala-Arg-Trp Chemical compound N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C12)C(=O)O DWINFPQUSSHSFS-UVBJJODRSA-N 0.000 description 1
- BEMGNWZECGIJOI-WDSKDSINSA-N Ala-Gly-Glu Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O BEMGNWZECGIJOI-WDSKDSINSA-N 0.000 description 1
- RMAWDDRDTRSZIR-ZLUOBGJFSA-N Ala-Ser-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O RMAWDDRDTRSZIR-ZLUOBGJFSA-N 0.000 description 1
- VRTOMXFZHGWHIJ-KZVJFYERSA-N Ala-Thr-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VRTOMXFZHGWHIJ-KZVJFYERSA-N 0.000 description 1
- JQFJNGVSGOUQDH-XIRDDKMYSA-N Arg-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCN=C(N)N)N)C(O)=O)=CNC2=C1 JQFJNGVSGOUQDH-XIRDDKMYSA-N 0.000 description 1
- JPAWCMXVNZPJLO-IHRRRGAJSA-N Arg-Ser-Phe Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JPAWCMXVNZPJLO-IHRRRGAJSA-N 0.000 description 1
- QTAIIXQCOPUNBQ-QXEWZRGKSA-N Arg-Val-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O QTAIIXQCOPUNBQ-QXEWZRGKSA-N 0.000 description 1
- SUMJNGAMIQSNGX-TUAOUCFPSA-N Arg-Val-Pro Chemical compound CC(C)[C@H](NC(=O)[C@@H](N)CCCNC(N)=N)C(=O)N1CCC[C@@H]1C(O)=O SUMJNGAMIQSNGX-TUAOUCFPSA-N 0.000 description 1
- NSTBNYOKCZKOMI-AVGNSLFASA-N Asn-Tyr-Glu Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O NSTBNYOKCZKOMI-AVGNSLFASA-N 0.000 description 1
- LRCIOEVFVGXZKB-BZSNNMDCSA-N Asn-Tyr-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O LRCIOEVFVGXZKB-BZSNNMDCSA-N 0.000 description 1
- VBVKSAFJPVXMFJ-CIUDSAMLSA-N Asp-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)O)N VBVKSAFJPVXMFJ-CIUDSAMLSA-N 0.000 description 1
- ICTXFVKYAGQURS-UBHSHLNASA-N Asp-Asn-Trp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O ICTXFVKYAGQURS-UBHSHLNASA-N 0.000 description 1
- DZQKLNLLWFQONU-LKXGYXEUSA-N Asp-Cys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC(=O)O)N)O DZQKLNLLWFQONU-LKXGYXEUSA-N 0.000 description 1
- SWTQDYFZVOJVLL-KKUMJFAQSA-N Asp-His-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC(=O)O)N)O SWTQDYFZVOJVLL-KKUMJFAQSA-N 0.000 description 1
- UKGGPJNBONZZCM-WDSKDSINSA-N Asp-Pro Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(O)=O UKGGPJNBONZZCM-WDSKDSINSA-N 0.000 description 1
- KGHLGJAXYSVNJP-WHFBIAKZSA-N Asp-Ser-Gly Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O KGHLGJAXYSVNJP-WHFBIAKZSA-N 0.000 description 1
- YIDFBWRHIYOYAA-LKXGYXEUSA-N Asp-Ser-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YIDFBWRHIYOYAA-LKXGYXEUSA-N 0.000 description 1
- RSMZEHCMIOKNMW-GSSVUCPTSA-N Asp-Thr-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O RSMZEHCMIOKNMW-GSSVUCPTSA-N 0.000 description 1
- USENATHVGFXRNO-SRVKXCTJSA-N Asp-Tyr-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(O)=O)C(O)=O)CC1=CC=C(O)C=C1 USENATHVGFXRNO-SRVKXCTJSA-N 0.000 description 1
- 101150049556 Bcr gene Proteins 0.000 description 1
- 208000025721 COVID-19 Diseases 0.000 description 1
- 101710132601 Capsid protein Proteins 0.000 description 1
- 101710094648 Coat protein Proteins 0.000 description 1
- 102100031673 Corneodesmosin Human genes 0.000 description 1
- 101710139375 Corneodesmosin Proteins 0.000 description 1
- JUNZLDGUJZIUCO-IHRRRGAJSA-N Cys-Pro-Tyr Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CS)N)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O JUNZLDGUJZIUCO-IHRRRGAJSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- CAXXTYYGFYTBPV-IUCAKERBSA-N Gln-Leu-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O CAXXTYYGFYTBPV-IUCAKERBSA-N 0.000 description 1
- HPBKQFJXDUVNQV-FHWLQOOXSA-N Gln-Tyr-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O HPBKQFJXDUVNQV-FHWLQOOXSA-N 0.000 description 1
- MLCPTRRNICEKIS-FXQIFTODSA-N Glu-Asn-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MLCPTRRNICEKIS-FXQIFTODSA-N 0.000 description 1
- RDDSZZJOKDVPAE-ACZMJKKPSA-N Glu-Asn-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O RDDSZZJOKDVPAE-ACZMJKKPSA-N 0.000 description 1
- IOUQWHIEQYQVFD-JYJNAYRXSA-N Glu-Leu-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IOUQWHIEQYQVFD-JYJNAYRXSA-N 0.000 description 1
- HQTDNEZTGZUWSY-XVKPBYJWSA-N Glu-Val-Gly Chemical compound CC(C)[C@H](NC(=O)[C@@H](N)CCC(O)=O)C(=O)NCC(O)=O HQTDNEZTGZUWSY-XVKPBYJWSA-N 0.000 description 1
- QSDKBRMVXSWAQE-BFHQHQDPSA-N Gly-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN QSDKBRMVXSWAQE-BFHQHQDPSA-N 0.000 description 1
- XBWMTPAIUQIWKA-BYULHYEWSA-N Gly-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CN XBWMTPAIUQIWKA-BYULHYEWSA-N 0.000 description 1
- COVXELOAORHTND-LSJOCFKGSA-N Gly-Ile-Val Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O COVXELOAORHTND-LSJOCFKGSA-N 0.000 description 1
- CVFOYJJOZYYEPE-KBPBESRZSA-N Gly-Lys-Tyr Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CVFOYJJOZYYEPE-KBPBESRZSA-N 0.000 description 1
- FFALDIDGPLUDKV-ZDLURKLDSA-N Gly-Thr-Ser Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O FFALDIDGPLUDKV-ZDLURKLDSA-N 0.000 description 1
- TVTZEOHWHUVYCG-KYNKHSRBSA-N Gly-Thr-Thr Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O TVTZEOHWHUVYCG-KYNKHSRBSA-N 0.000 description 1
- FOKISINOENBSDM-WLTAIBSBSA-N Gly-Thr-Tyr Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O FOKISINOENBSDM-WLTAIBSBSA-N 0.000 description 1
- UMBDRSMLCUYIRI-DVJZZOLTSA-N Gly-Trp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)CN)O UMBDRSMLCUYIRI-DVJZZOLTSA-N 0.000 description 1
- GWNIGUKSRJBIHX-STQMWFEESA-N Gly-Tyr-Arg Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)CN)O GWNIGUKSRJBIHX-STQMWFEESA-N 0.000 description 1
- DNAZKGFYFRGZIH-QWRGUYRKSA-N Gly-Tyr-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 DNAZKGFYFRGZIH-QWRGUYRKSA-N 0.000 description 1
- KSOBNUBCYHGUKH-UWVGGRQHSA-N Gly-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN KSOBNUBCYHGUKH-UWVGGRQHSA-N 0.000 description 1
- 102100021181 Golgi phosphoprotein 3 Human genes 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- VOEGKUNRHYKYSU-XVYDVKMFSA-N His-Asp-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O VOEGKUNRHYKYSU-XVYDVKMFSA-N 0.000 description 1
- OEROYDLRVAYIMQ-YUMQZZPRSA-N His-Gly-Asp Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O OEROYDLRVAYIMQ-YUMQZZPRSA-N 0.000 description 1
- RLAOTFTXBFQJDV-KKUMJFAQSA-N His-Phe-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CN=CN1 RLAOTFTXBFQJDV-KKUMJFAQSA-N 0.000 description 1
- HIJIJPFILYPTFR-ACRUOGEOSA-N His-Tyr-Tyr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O HIJIJPFILYPTFR-ACRUOGEOSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- IIWQTXMUALXGOV-PCBIJLKTSA-N Ile-Phe-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)O)C(=O)O)N IIWQTXMUALXGOV-PCBIJLKTSA-N 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 1
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 1
- QLQHWWCSCLZUMA-KKUMJFAQSA-N Leu-Asp-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QLQHWWCSCLZUMA-KKUMJFAQSA-N 0.000 description 1
- QCSFMCFHVGTLFF-NHCYSSNCSA-N Leu-Asp-Val Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O QCSFMCFHVGTLFF-NHCYSSNCSA-N 0.000 description 1
- BDFHWFUAQLIMJO-KXNHARMFSA-N Lys-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCCN)N)O BDFHWFUAQLIMJO-KXNHARMFSA-N 0.000 description 1
- 101710125418 Major capsid protein Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- ZDJICAUBMUKVEJ-CIUDSAMLSA-N Met-Ser-Gln Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(N)=O ZDJICAUBMUKVEJ-CIUDSAMLSA-N 0.000 description 1
- ATBJCCFCJXCNGZ-UFYCRDLUSA-N Met-Tyr-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 ATBJCCFCJXCNGZ-UFYCRDLUSA-N 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- 101710141454 Nucleoprotein Proteins 0.000 description 1
- DJPXNKUDJKGQEE-BZSNNMDCSA-N Phe-Asp-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O DJPXNKUDJKGQEE-BZSNNMDCSA-N 0.000 description 1
- OJUMUUXGSXUZJZ-SRVKXCTJSA-N Phe-Asp-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O OJUMUUXGSXUZJZ-SRVKXCTJSA-N 0.000 description 1
- FRPVPGRXUKFEQE-YDHLFZDLSA-N Phe-Asp-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O FRPVPGRXUKFEQE-YDHLFZDLSA-N 0.000 description 1
- BPIFSOUEUYDJRM-DCPHZVHLSA-N Phe-Trp-Ala Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](C)C(O)=O)C1=CC=CC=C1 BPIFSOUEUYDJRM-DCPHZVHLSA-N 0.000 description 1
- JLDZQPPLTJTJLE-IHPCNDPISA-N Phe-Trp-Asp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)N[C@@H](CC(=O)O)C(=O)O)N JLDZQPPLTJTJLE-IHPCNDPISA-N 0.000 description 1
- WDOCBGZHAQQIBL-IHPCNDPISA-N Phe-Trp-Ser Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CO)C(O)=O)C1=CC=CC=C1 WDOCBGZHAQQIBL-IHPCNDPISA-N 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- SSWJYJHXQOYTSP-SRVKXCTJSA-N Pro-His-Gln Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(N)=O)C(O)=O SSWJYJHXQOYTSP-SRVKXCTJSA-N 0.000 description 1
- DRKAXLDECUGLFE-ULQDDVLXSA-N Pro-Leu-Phe Chemical compound CC(C)C[C@H](NC(=O)[C@@H]1CCCN1)C(=O)N[C@@H](Cc1ccccc1)C(O)=O DRKAXLDECUGLFE-ULQDDVLXSA-N 0.000 description 1
- AWQGDZBKQTYNMN-IHRRRGAJSA-N Pro-Phe-Asp Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N[C@@H](CC(=O)O)C(=O)O AWQGDZBKQTYNMN-IHRRRGAJSA-N 0.000 description 1
- HOTVCUAVDQHUDB-UFYCRDLUSA-N Pro-Phe-Tyr Chemical compound C([C@@H](C(=O)O)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H]1NCCC1)C1=CC=C(O)C=C1 HOTVCUAVDQHUDB-UFYCRDLUSA-N 0.000 description 1
- 101710083689 Probable capsid protein Proteins 0.000 description 1
- 108010054530 RGDN peptide Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- UJTVWLNMFSYGTA-UHFFFAOYSA-N Ser Gly Trp Tyr Chemical compound C=1NC2=CC=CC=C2C=1CC(NC(=O)CNC(=O)C(CO)N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 UJTVWLNMFSYGTA-UHFFFAOYSA-N 0.000 description 1
- UBRXAVQWXOWRSJ-ZLUOBGJFSA-N Ser-Asn-Asp Chemical compound C([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CO)N)C(=O)N UBRXAVQWXOWRSJ-ZLUOBGJFSA-N 0.000 description 1
- MUARUIBTKQJKFY-WHFBIAKZSA-N Ser-Gly-Asp Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O MUARUIBTKQJKFY-WHFBIAKZSA-N 0.000 description 1
- MIJWOJAXARLEHA-WDSKDSINSA-N Ser-Gly-Glu Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O MIJWOJAXARLEHA-WDSKDSINSA-N 0.000 description 1
- UGTZYIPOBYXWRW-SRVKXCTJSA-N Ser-Phe-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O UGTZYIPOBYXWRW-SRVKXCTJSA-N 0.000 description 1
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 description 1
- SQHKXWODKJDZRC-LKXGYXEUSA-N Ser-Thr-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O SQHKXWODKJDZRC-LKXGYXEUSA-N 0.000 description 1
- JMGJDTNUMAZNLX-RWRJDSDZSA-N Thr-Glu-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JMGJDTNUMAZNLX-RWRJDSDZSA-N 0.000 description 1
- XPNSAQMEAVSQRD-FBCQKBJTSA-N Thr-Gly-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)NCC(O)=O XPNSAQMEAVSQRD-FBCQKBJTSA-N 0.000 description 1
- QQWNRERCGGZOKG-WEDXCCLWSA-N Thr-Gly-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O QQWNRERCGGZOKG-WEDXCCLWSA-N 0.000 description 1
- JQAWYCUUFIMTHE-WLTAIBSBSA-N Thr-Gly-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JQAWYCUUFIMTHE-WLTAIBSBSA-N 0.000 description 1
- MXDOAJQRJBMGMO-FJXKBIBVSA-N Thr-Pro-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O MXDOAJQRJBMGMO-FJXKBIBVSA-N 0.000 description 1
- YRJOLUDFVAUXLI-GSSVUCPTSA-N Thr-Thr-Asp Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(O)=O YRJOLUDFVAUXLI-GSSVUCPTSA-N 0.000 description 1
- OFNPHOGOJLNVLL-KCTSRDHCSA-N Trp-Ala-His Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N OFNPHOGOJLNVLL-KCTSRDHCSA-N 0.000 description 1
- SCQBNMKLZVCXNX-ZFWWWQNUSA-N Trp-Arg-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(=O)O)N SCQBNMKLZVCXNX-ZFWWWQNUSA-N 0.000 description 1
- OGZRZMJASKKMJZ-XIRDDKMYSA-N Trp-Leu-Asp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N OGZRZMJASKKMJZ-XIRDDKMYSA-N 0.000 description 1
- GQNCRIFNDVFRNF-BPUTZDHNSA-N Trp-Pro-Asp Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O GQNCRIFNDVFRNF-BPUTZDHNSA-N 0.000 description 1
- IIJWXEUNETVJPV-IHRRRGAJSA-N Tyr-Arg-Ser Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N)O IIJWXEUNETVJPV-IHRRRGAJSA-N 0.000 description 1
- GFHYISDTIWZUSU-QWRGUYRKSA-N Tyr-Asn-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O GFHYISDTIWZUSU-QWRGUYRKSA-N 0.000 description 1
- MVYRJYISVJWKSX-KBPBESRZSA-N Tyr-His-Gly Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)NCC(=O)O)N)O MVYRJYISVJWKSX-KBPBESRZSA-N 0.000 description 1
- KIJLSRYAUGGZIN-CFMVVWHZSA-N Tyr-Ile-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O KIJLSRYAUGGZIN-CFMVVWHZSA-N 0.000 description 1
- MDXLPNRXCFOBTL-BZSNNMDCSA-N Tyr-Ser-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O MDXLPNRXCFOBTL-BZSNNMDCSA-N 0.000 description 1
- ABZWHLRQBSBPTO-RNXOBYDBSA-N Tyr-Trp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)[C@H](CC4=CC=C(C=C4)O)N ABZWHLRQBSBPTO-RNXOBYDBSA-N 0.000 description 1
- 108010064997 VPY tripeptide Proteins 0.000 description 1
- LTFLDDDGWOVIHY-NAKRPEOUSA-N Val-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H](C(C)C)N LTFLDDDGWOVIHY-NAKRPEOUSA-N 0.000 description 1
- KKHRWGYHBZORMQ-NHCYSSNCSA-N Val-Arg-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N KKHRWGYHBZORMQ-NHCYSSNCSA-N 0.000 description 1
- OVLIFGQSBSNGHY-KKHAAJSZSA-N Val-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C(C)C)N)O OVLIFGQSBSNGHY-KKHAAJSZSA-N 0.000 description 1
- KTEZUXISLQTDDQ-NHCYSSNCSA-N Val-Lys-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)O)C(=O)O)N KTEZUXISLQTDDQ-NHCYSSNCSA-N 0.000 description 1
- XPKCFQZDQGVJCX-RHYQMDGZSA-N Val-Lys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C(C)C)N)O XPKCFQZDQGVJCX-RHYQMDGZSA-N 0.000 description 1
- WMRWZYSRQUORHJ-YDHLFZDLSA-N Val-Phe-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)O)C(=O)O)N WMRWZYSRQUORHJ-YDHLFZDLSA-N 0.000 description 1
- DLRZGNXCXUGIDG-KKHAAJSZSA-N Val-Thr-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N)O DLRZGNXCXUGIDG-KKHAAJSZSA-N 0.000 description 1
- JSOXWWFKRJKTMT-WOPDTQHZSA-N Val-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N JSOXWWFKRJKTMT-WOPDTQHZSA-N 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 108010072041 arginyl-glycyl-aspartic acid Proteins 0.000 description 1
- 108010009111 arginyl-glycyl-glutamic acid Proteins 0.000 description 1
- 108010066119 arginyl-leucyl-aspartyl-serine Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010805 cDNA synthesis kit Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 239000003517 fume Substances 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 108010008237 glutamyl-valyl-glycine Proteins 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 108010036413 histidylglycine Proteins 0.000 description 1
- 108010092114 histidylphenylalanine Proteins 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 108010078274 isoleucylvaline Proteins 0.000 description 1
- 108010083708 leucyl-aspartyl-valine Proteins 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 238000002824 mRNA display Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108010053725 prolylvaline Proteins 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 238000002702 ribosome display Methods 0.000 description 1
- 238000007423 screening assay Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 210000004989 spleen cell Anatomy 0.000 description 1
- 230000009258 tissue cross reactivity Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011830 transgenic mouse model Methods 0.000 description 1
- 108010029384 tryptophyl-histidine Proteins 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 108010020532 tyrosyl-proline Proteins 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
-
- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
- G16B20/00—ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations
- G16B20/30—Detection of binding sites or motifs
-
- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
- G16B30/00—ICT specially adapted for sequence analysis involving nucleotides or amino acids
- G16B30/10—Sequence alignment; Homology search
-
- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
- G16B40/00—ICT specially adapted for biostatistics; ICT specially adapted for bioinformatics-related machine learning or data mining, e.g. knowledge discovery or pattern finding
- G16B40/10—Signal processing, e.g. from mass spectrometry [MS] or from PCR
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Medical Informatics (AREA)
- Analytical Chemistry (AREA)
- Theoretical Computer Science (AREA)
- Bioinformatics & Computational Biology (AREA)
- Genetics & Genomics (AREA)
- Evolutionary Biology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Data Mining & Analysis (AREA)
- Databases & Information Systems (AREA)
- Epidemiology (AREA)
- Evolutionary Computation (AREA)
- Computer Vision & Pattern Recognition (AREA)
- Public Health (AREA)
- Software Systems (AREA)
- Bioethics (AREA)
- Signal Processing (AREA)
- Artificial Intelligence (AREA)
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明公开了一种筛选特异BCR/TCR的方法及其系统。该方法包括以下步骤:筛选特异性BCR/TCR的方法,包括以下步骤:(1)提取免疫后的待测样本淋巴细胞中的mRNA;(2)逆转录处理步骤(1)所得mRNA后,再对其BCR/TCR可变区基因进行多重PCR扩增,然后进行高通量测序,并确定BCR/TCR可变区的核酸及氨基酸序列;(3)将测序得到的序列与未经抗原刺激样本的淋巴细胞BCR/TCR测序结果对比,获得相应的特异性免疫序列。本发明通过免疫图谱大数据进行对比,同时比较巨大数量的BCR/TCR序列,具有更高的特异性和准确性,可以极大提高抗体及T细胞受体筛选鉴定的效率,且筛选成本低。
Description
技术领域
本发明属于分子生物学和细胞免疫学技术领域,具体涉及一种筛选特异性抗体(BCR)及T细胞受体(TCR)的方法及其系统。
背景技术
单克隆抗体,是由单一B细胞产生的高度均一、识别特定抗原表位的抗体。单克隆抗体在医学与生物学基础研究,以及临床疾病的诊断、预防和治疗中都有广泛的应用前景。要得到针对特定抗原的单克隆抗体,需要进行抗体筛选。现有比较成熟的单克隆抗体筛选技术有:
1、杂交瘤技术。
将抗原免疫后的小鼠脾细胞与小鼠骨髓瘤细胞进行融合,形成的杂交细胞既可产生抗体,又可无限增殖。将融合的杂交细胞分成单细胞接种培养,通过酶联免疫吸附测定(ELISA)等方法筛选鉴定增殖后细胞分泌的抗体,找到能产生针对抗原具有高滴度抗体的细胞株,再从其中克隆出该抗体的基因序列。
杂交瘤技术的主要缺点是,后期鉴定过程需要进行大量重复性的培养、分离、鉴定实验,带来的是高昂的时间和物质成本。
2、噬菌体展示筛选技术。
将B淋巴细胞的抗体重链可变区(VH)和轻链可变区(VL)采用PCR方法进行扩增,将扩增的片段插入到噬菌体载体中,抗体分子以Fab片段或者单链抗体(scFv)的形式,与单链噬菌体外壳蛋白形成融合蛋白,从而展示在噬菌体表面。之后将噬菌体转染至宿主细胞中进行增殖,待成熟后释放出来,最后利用抗原筛选的方法经过亲和吸附、洗脱、扩增等步骤后即可获得靶抗原特异性的噬菌体及对应单克隆抗体。
为了得到更接近人源抗体的抗体进行筛选,还有酵母表面展示技术、核糖体展示与mRNA展示筛选技术等,其方法与噬菌体展示筛选类似。各种展示技术筛选抗体的缺点与杂交瘤技术类似,后期鉴定过程工作量大,时间和物质成本高。
3、单细胞克隆技术。
先通过流式细胞分析等方法,筛选出表面受体能识别特定抗原的B淋巴细胞或T淋巴细胞。再直接从单个B细胞或T细胞中,克隆出它携带的B细胞受体(BCR,分泌到体外即为抗体)或T细胞受体(TCR)基因。
单克隆技术的缺点有两方面:一方面,单个淋巴细胞表面的BCR或TCR数量有限,这就限制了利用抗原分选单细胞的灵敏度和特异性,分选出的细胞不能确保必然是阳性细胞,后期仍然需要对得到的抗体/TCR进行比较多的鉴定;另一方面,单细胞克隆的难度较高,对细胞的质量、操作人员的技能、试剂的品质都有很高要求。
现有单克隆抗体筛选技术还有三个显著的不足:第一是对于人源抗体的筛选比较困难,因为无法像免疫动物一样使用抗原和佐剂对人体进行反复免疫。如果使用转基因小鼠(携带人源免疫球蛋白基因),则成本会更高。这就使得制备人源抗体要么需要在筛选鉴定中投入更多资源,要么需要对动物抗体进行人源化。
第二是对针对复杂抗原制备单克隆抗体存在困难。当需要针对较大较复杂的抗原(如膜蛋白复合物、肿瘤细胞等)制备抗体时,选择制备用于免疫的抗原本身就有较大难度且存在失败风险,而免疫后也不能保证得到功能上满足要求的抗体(如选择多肽段免疫所得抗体,未必是中和抗体)。如果同时选取多种抗原进行免疫和筛选,则会显著提高单克隆抗体制备的成本。
第三,需要确定抗原进行筛选鉴定
基于高通量测序技术和免疫图谱大数据库,我们建立了一种新型的单克隆抗体筛选技术。通过将被特定抗原免疫的实验动物或受试者的免疫图谱,与对照组免疫图谱大数据库进行比较和分析,利用特定算法筛选出最有可能针对特定抗原的抗体或TCR基因序列,可以极大降低后期鉴定的工作量,提高单克隆抗体的筛选效率。
发明内容
针对现有技术中的上述不足,本发明提供一种筛选特异性BCR/TCR的方法及其系统,本发明通过免疫图谱大数据进行对比,同时比较巨大数量的BCR/TCR序列,具有更高的特异性和准确性,可以极大提高单克隆抗体筛选鉴定的效率。
为实现上述目的,本发明解决其技术问题所采用的技术方案是:
一种筛选特异性BCR/TCR的方法,包括以下步骤:
(1)分别提取N个免疫后待测样本的淋巴细胞中的mRNA;
(2)逆转录处理步骤(1)所得mRNA后,再对其BCR/TCR可变区基因进行多重PCR扩增,然后进行高通量测序,并确定BCR/TCR可变区的核酸及氨基酸序列;
(3)将测序得到的序列与未经抗原刺激样本的淋巴细胞BCR/TCR测序结果对比,获得相应的特异性免疫序列。
进一步地,待测样本为包含淋巴细胞的外周血、脾脏组织、肿瘤组织或体外培养的淋巴细胞。
进一步地,待测样本为人源样本或动物源样本。
进一步地,步骤(3)还包括:将所有免疫后样本BCR/TCR序列归总去重,再去除所有与未免疫样本测序结果序列集中所包含序列相重复的序列,建立特征性BCR/TCR序列集。
进一步地,特征性BCR/TCR序列集中特异性免疫序列的具体获得方式为:
(1)从特征性BCR/TCR序列集中去除只在<N×10%个免疫后样本中出现的序列;
(2)对特征性BCR/TCR序列集中所含有的共M种具有唯一性的BCR/TCR CDR3序列分别在N个免疫后样本测序结果中出现的次数进行统计,计为X1,1、X1,2、X1,3……X1,M,X2,1、X2,2、X2,3……X2,M,……,XN,1、XN,2、XN,3……XN,M;
(3)将特征性序列集中相同BCR/TCR CDR3序列在N个免疫后样本测序结果中出现次数求和,第j种CDR3序列出现次数之和为:
(4)将特征性序列集中所含有的共M种BCR/TCR CDR3序列按Yj值从高至低排序,排序前100个或前M×10%个序列即为与抗原相对应的特异性BCR/TCR CDR3序列。
进一步地,BCR序列的选择还可以参考抗原的电荷、疏水性等生化特性。
进一步地,筛选新型冠状病毒COVID-19感染后外周血中的特征性的抗体重链序列的方法,包括以下步骤:
(1)分别采集健康人和未感染新冠的其它疾病病人,以及新型冠状病毒感染患者的外周血,并提取外周血淋巴细胞中的mRNA;
(2)逆转录处理步骤(1)所得mRNA后,再对其抗体重链可变区基因进行多重PCR扩增,然后进行高通量测序,并确定抗体重链可变区的核酸及氨基酸序列,尽量保证每个样本的功能性抗体重链序列总数综合不低于30000,并建立抗体序列集;
(3)对每个样本的功能性抗体重链序列进行随机不放回抽样,使每个样本的功能性抗体重链序列数量总和为30000,此功能性抗体的集合定义为该样本的抗体重链序列集。如某样本功能性抗体重链序列数量不足30000,则使用全部测序所得功能性抗体重链序列为该样本的序列集。
(4)将健康人和未感染新冠的其它疾病病人作为对照组,并对其所有的测序所得功能性抗体重链序列归总去重,设为对照序列集;
(5)将N个新型冠状病毒感染患者的所有抗体重链序列归总去重,再去除所有与对照序列集中所包含序列相重复的序列,即可得到相应的新型冠状病毒COVID-19感染后外周血中的特征性抗体重链序列集。
(6)从新冠特征性抗体重链序列集中去除只在<N×10%个新冠病人样本中出现的序列。
(7)对新冠特征性抗体重链序列集中所含有的共M种具有唯一性的抗体重链CDR3序列分别在N个新冠病人样本测序结果中出现的次数进行统计,计为X1,1、X1,2、X1,3……X1,M,X2,1、X2,2、X2,3……X2,M,……,XN,1、XN,2、XN,3……XN,M;
(8)将新冠特征性抗体序列集中相同抗体重链CDR3序列在N个新冠病人样本测序结果中出现次数求和,第j种CDR3序列出现次数之和为:
(9)将新冠特征性序列集中所含有的共M种抗体重链CDR3序列按Yj值从高至低排序,排序前100个或前M×10%个序列即为与新冠病毒抗原相对应的特异性抗体重链CDR3序列。
(10)参考新冠病毒抗原的电荷、疏水性等生化特性,对抗体重链CDR3序列进行筛选。
进一步地,新型冠状病毒特异性抗体重链CDR3氨基酸序列如SEQ ID NO.1~39所示。
一种用于筛选特异性BCR/TCR的系统,该系统包括测序单元,测序单元用于对免疫后样本的淋巴细胞的BCR/TCR可变区进行序列测定,以便获得测序数据;
序列集建立单元,序列集建立单元用于对所有的BCR/TCR序列数量进行检测,并根据检测结果建立相应序列集;
序列比对单元,该单元将免疫后样本建立的序列集与对照组建立的序列集进行比对去重,并获得相应的特征性BCR/TCR序列集。
该系统能够采用上述方法对特征性BCR/TCR序列集进行筛选,得到特异性BCR/TCR序列。
本发明的有益效果为:
1、本发明通过免疫图谱大数据进行对比,同时比较巨大数量的BCR/TCR序列,具有更高的特异性和准确性,可以极大提高单克隆抗体筛选鉴定的效率。
2、高通量测序成本已经降到很低水平;测序样品处理的人力成本,也大大低于传统抗体筛选技术后期所需大量鉴定工作。
3、本发明可以使用包含淋巴细胞的任何样本,包括且不限于:外周血、脾脏组织、肿瘤组织、体外培养的淋巴细胞。样本可以来自动物,也可以来自人体。可以是主动免疫,也可以是来自患者。
4、可以一次得到针对复杂抗原的大量备选BCR/TCR,经过较简单鉴定后,即可用于后续研究甚至临床试验。
附图说明
图1为新型冠状病毒特征抗体重链序列筛选对照图。
具体实施方式
下面对本发明的具体实施方式进行描述,以便于本技术领域的技术人员理解本发明,但应该清楚,本发明不限于具体实施方式的范围,对本技术领域的普通技术人员来讲,只要各种变化在所附的权利要求限定和确定的本发明的精神和范围内,这些变化是显而易见的,一切利用本发明构思的发明创造均在保护之列。
实施例1通过高通量测序得到新型冠状病毒(COVID-19)感染后外周血中特征性的抗体重链序列集
1、采集1377名对照组(包括健康人和未感染新冠的病人)、94名新型冠状病毒感染患者的外周血:
(1)使用EDTA无菌采血管(不要用肝素或枸橼酸),采血0.5-1mL,轻弹采血管使血液与抗凝剂混匀。采集好的血液如来不及处理,可以暂时放在4℃冰箱保存(不超过2小时)。
(2)竖立采血管,使用一支全新的一次性注射器,针头穿过橡胶塞(刚穿过即可,不要打开采血管盖子)向采血管内注入5mL TRIzol(TRIsol,RNAiso plus,SuperfecTRI都可以)。如血液经过4℃冰箱保存已出现分层,注入TRIzol前请摇晃采血管,使血液重新混合均匀。
如采血管已失去真空,注入TRIzol过程中感觉明显阻力,可缓缓回拉注射器活塞释放管顶空气压力,再继续注入。注入完成后,也应缓缓回拉注射器活塞释放管顶空气压力,以免后续步骤内部液体从针孔喷出。使用后的一次性注射器针头必须与采血针一起妥善处理。
(3)反复颠倒摇晃采血管,使TRIzol与血液混匀,直至看不到明显血块。室温静置5min,使细胞充分裂解,再颠倒混匀。步骤2-3应在洁净通风处完成,建议使用生物安全柜/超净台/通风橱。
(4)裂解好的外周血样本可以保存在-20℃或-80℃冰箱,或冷链寄送。
2、淋巴细胞基因组mRNA的提取及检测
(1)取1mL血液裂解物,加入200μL氯仿,手动剧烈摇晃15s后,室温静置5min。
(2)4℃,12000g离心15min,吸取上层水相,转移至另一离心管,约450μL。
(3)加入500μL异丙醇混匀,室温静置10min。
(4)4℃,12000g离心10min,弃上清,RNA沉于管底。
(5)加入1mL 75%乙醇(DEPC水配制),温和震荡离心管,使RNA沉淀整片悬浮。
(6)4℃,7500g离心5min,弃上清,室温放置5-8min,挥发酒精,不能过于干燥。
(7)用30μL DEPC/无酶水溶解RNA沉淀,少量分装后,做好标记,-80℃冰箱保存。
(8)取2μL RNA,用微量紫外分光光度计检测浓度和OD值,取2μL RNA电泳检测条带,使用2%的凝胶,DL2000Marker,同时立刻进行反转录实验。
检测结果为RNA总量>5μg,体积>20μL,OD260/280比值1.8以上,同时2%琼脂糖电泳检测,出现三条带,为28sRNA、18sRNA、5sRNA,其中28S为18S条带亮度的1~2倍,5S条带最浅。
3、逆转录及PCR扩增
(1)逆转录反应
使用同科公司试剂盒(TonkBioTM First chain cDNA synthesis kit)进行逆转录反应,反应体系和反应条件如表1所示:
表1 逆转录反应体系和反应条件
其中,BRTmix引物的名称和序列如表2所示,引物浓度1:1:1混合,总浓度为20μM。
表2 BRTmix引物
BRTmix | 序列5’-3’ |
hIgM-RT | AGGAAGTCCTGTGCGAGGCA |
hIgG-RT | CGGGGAAGTAGTCCTTGACC |
hIgA-RT | CGCTCCAGGTCACACTGAGT |
(2)第一轮PCR:
对cDNA的BCR基因进行特异性扩增反应,其中所用的PCR酶为同科公司Taq酶(货号TK01015),反应体系及条件如表3和表4所示:
表3 第一轮PCR反应体系
逆转录产物 | 2μL |
VHmix引物(20uM) | 0.5μL |
JHmix(10uM) | 1μL |
10×Buffer | 2μL |
Mgcl2(25mM) | 1.6μL |
dNTP(10mM each) | 0.8μL |
Taq酶 | 1.2μL |
ddH2O | 10.9μL |
total | 20μL |
表4 PCR程序
其中,VHmix引物为5’端多重PCR引物,引物名称和序列见表5,按1:1混合,总浓度为20μM。
表5 VHmix引物
VHmix | 序列5’-3’ |
5VH1a | CAGGTGCAGCTGGTGCAGTCTGGGG |
5VH1b | CAGGTCCAGCTTGTGCAGTCTGGGG |
5VH1C | CAGGTTCAGCTGGTGCAGTCTGGAG |
5VH2 | CAGRTCACCTTGARGGAGTCTGGTCCT |
5VH3a | GAGGTGCAGCTGGTGGAGTCTGGGG |
5VH3b | CAGGTGCAGCTGGTGGAGTCTGGGG |
VH3-23 | GAGGTGCAGCTGTTGGAGTCTGGGG |
VH3-74 | GAGGTGCAGCTGGTGGAGTCCGGGG |
5VH4 | CAGGTGCAGCTGCAGGAGTCGGGC |
5VH4-34 | CAGGTGCAGCTACAGCAGTGGGGC |
5VH4-39 | CAGCTGCAGCTGCAGGAGTCGGGC |
5VH5 | GARGTGCAGCTGGTGCAGTCTGGAG |
5VH6 | CAGGTACAGCTGCAGCAGTCAGGTCC |
5VH7 | CAGGTGCAGCTGGTGCAATCTGGGTC |
JHmix引物为3’端多重PCR引物,引物名称和序列见表6,按1:1混合,总浓度为10uM,其中小写字母部分是与第二轮3’端引物barcode的保护序列配对。
表6 JHmix引物
第一轮PCR结束后,不进行电泳检测,直接进行第二轮PCR扩增。
(3)第二轮PCR:
使用同科公司Taq酶(货号TK01015)进行第二轮PCR反应,反应体系和反应条件如表7和表8所示:
表7 第二轮PCR反应体系
第一轮PCR产物 | 2μL |
VHmix引物(20uM) | 1μL |
JHxBCX(20uM) | 1μL |
10×Buffer | 2μL |
Mgcl2(25mM) | 1.6μL |
dNTP(10mM each) | 0.8μL |
酶 | 1.2μL |
ddH2O | 10.4μL |
total | 20μL |
表8 PCR程序
其中VHmix引物为5’端多重PCR引物,即第一轮PCR中的VHmix。
JHxBCX为3’端引物,引物名称和序列见表9,包含有10个不同barcode的引物序列,引物序列包括barcode序列(下划线部分)、barcode保护识别序列(小写字母部分)和特异性扩增序列,一个样品选用一种JHxBCX引物,对应唯一一个barcode,以此根据barcode序列区分不同的样本。
表9 JHxBCX引物
(4)电泳检测扩增效果,目的条带在400多bp处。
(5)磁珠纯化:一个样本做1~2管第二轮PCR后即可纯化。在PCR产物中加入等体积的DNA片段分选纯化磁珠(百迈格生物货号BMSX),使用30μL水洗脱。使用微量分光光度计检测纯化后产物浓度。
(6)等浓度混合不同barcode样本,建库测序及后续分析。
4、建库、测序及数据分析获得免疫图谱
(1)、建库及高通量测序
a)建库
严格按照操作手册操作,加上测序接头及定量工作。
根据试剂盒说明书进行建库,包括Ion Plus Fragment Library Kit(货号4471252)、Ion Xpress Barcode Adaptors 1-16Kit(货号4471250)、Agencourt AMPure XP(货号A63881),仪器包括PCR仪、Qubit定量仪及核酸定量试剂。
b)模板制备及上机测序
Ion chef-自动化模板制备仪,安装好试剂及耗材即可自动进行实验,完成文库加上ISP磁珠、富集、loading芯片工作。
Ion S5-测序仪,完成测序工作及初步数据处理。
使用试剂包括Ion 520/530 ExT-Chef-4rxns&4 Init NEW-For 600bp(货号A30670)、Ion 530 Chip Kit(货号A27764)。
(2)数据分析获得免疫图谱
高通量测序完成后,使用FQ文件进行数据分析,序列进行IgBLAST,通过分析抗体重链基因的V、D和J基因,确定抗体重链的核酸及氨基酸序列,尽量保证每个样本的功能性抗体重链序列总数综合不低于30000。
(3)对每个样本的功能性抗体重链序列进行随机不放回抽样,使每个样本的功能性抗体重链序列数量总和为30000,此功能性抗体重链的集合定义为该样本的抗体重链序列集。如某样本功能性抗体重链序列数量不足30000,则使用全部测序所得功能性抗体重链序列为该样本的抗体重链序列集;
(4)通过分析抗体重链测序数据,确定抗新型冠状病毒特征抗体重链序列集:
a)将1377名对照组样本的所有抗体重链序列归总去重,设为对照序列集;
b)将94名新型冠状病毒感染样本的所有抗体重链序列归总去重,再去除所有与对照序列集中包含序列重复的序列,设为新型冠状病毒特征抗体序列集。如图1所示,其中横坐标代表某一特定氨基酸组合的抗体重链序列被加入对照序列集合或新型冠状病毒感染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的对数值。
实施例2、从新型冠状病毒感染后外周血中特征性的抗体重链序列中筛选出最具潜力的新冠抗体重链序列
1、从新型冠状病毒特征抗体重链序列集中,去除所有出现在不到10个新型冠状病毒感染者样本(<10%新冠感染病人人数)里的抗体重链序列。本步骤共选出77条抗体CDR3序列。
2、将步骤1筛选后的抗体重链序列,按“任一唯一性的抗体重链CDR3序列在所有新型冠状病毒感染病人样本中重复出现次数的总和”从高到低排序,选择排名前100或前10%者。因步骤1所选出序列较少,本步骤全部保留。
3、根据抗原蛋白特性,筛选抗体重链CDR3序列:因为新冠病毒S蛋白受体结合区域(RBD)含有较多正电荷,因此在步骤2选择的序列中,选择抗体重链CDR3序列中含有较多(1个及以上)净负电荷的序列。本步骤共选出39条序列,即将其为新型冠状病毒特异抗体CDR3序列,其氨基酸序列如表10中SEQ ID NO.1~39所示。
表10 新冠特异性抗体CDR3序列
序列表
<110> 成都益安博生物技术有限公司
<120> 一种筛选特异BCR/TCR的方法及其系统
<160> 39
<170> SIPOSequenceListing 1.0
<210> 1
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 1
Ala Arg Glu Pro Leu Phe Asp Thr Thr Asp His Tyr Val Phe Asp Gln
1 5 10 15
<210> 2
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 2
Ala Lys Asp Pro Phe Tyr Asp Phe Trp Ser Gly Tyr Tyr Phe Asp Tyr
1 5 10 15
<210> 3
<211> 12
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 3
Val Arg Glu Thr Pro Gly Ala Gly Glu Phe Asp Pro
1 5 10
<210> 4
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 4
Ala Ser Asp Arg Glu Trp Gln Leu Gly Thr Gly Gly Trp Leu Asp Pro
1 5 10 15
<210> 5
<211> 13
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 5
Ala Arg Glu Ser Asn Asp Tyr Gly Ser Pro Phe Asp Tyr
1 5 10
<210> 6
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 6
Ala Arg Glu Tyr Leu Glu Trp Phe Asp Arg Tyr Tyr Met Asp Val
1 5 10 15
<210> 7
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 7
Val Lys Asp Met Ser Gln Thr Glu Ile Leu Tyr Tyr Phe Asp Phe
1 5 10 15
<210> 8
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 8
Ala Lys Glu Ser Gly Phe Tyr Tyr Glu Asn Ser Gly Tyr Leu Asp Ser
1 5 10 15
<210> 9
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 9
Ala Lys Glu Ser Gly Phe Tyr Tyr Glu Asn Ser Gly Tyr Leu Asp Ser
1 5 10 15
<210> 10
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 10
Ala Lys Asp Pro His Tyr Asp Phe Trp Ser Gly Tyr Tyr Phe Asp Tyr
1 5 10 15
<210> 11
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 11
Ala Lys Asp Pro His Tyr Asp Phe Trp Ser Gly Tyr Tyr Phe Asp Tyr
1 5 10 15
<210> 12
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 12
Ala Thr Asp Arg Pro Ile Asp Val Val Ile Tyr Ala Leu Asp Phe
1 5 10 15
<210> 13
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 13
Ala Lys Asp Lys Thr Pro Asp Asn His Phe Trp Asp Tyr Phe Asp Tyr
1 5 10 15
<210> 14
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 14
Thr Thr Asp Ser Ser Gly Trp Arg Gly Asp Asn Trp Phe Asp Pro
1 5 10 15
<210> 15
<211> 13
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 15
Ala Arg Asp Leu Ser Phe Gly Ser Leu His Phe Ala Asp
1 5 10
<210> 16
<211> 14
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 16
Ala Arg Glu Gly Ile Val Gly Ala Thr Thr Gly Leu Asp Tyr
1 5 10
<210> 17
<211> 12
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 17
Ala Arg Trp His Gly Asp Phe Trp Ala Phe Asp Ser
1 5 10
<210> 18
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 18
Ala Arg Gly Glu Leu Tyr Met Tyr Phe Tyr Asn Gly Met Asp Val
1 5 10 15
<210> 19
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 19
Ala Thr Arg Trp Pro Asp Gln Tyr Tyr Tyr Tyr Gly Met Asp Val
1 5 10 15
<210> 20
<211> 18
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 20
Ala Arg His Gly Val Val Val Asp Thr His Tyr Tyr Asn Tyr Tyr Met
1 5 10 15
Asp Val
<210> 21
<211> 19
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 21
Ala Arg His Trp Ala His Asp Cys Thr Ser Thr Asn Cys Pro Tyr Tyr
1 5 10 15
Met Asp Val
<210> 22
<211> 18
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 22
Ala Arg Gly Gly Asp Ile Val Val Val Pro Val Ala Ile Tyr Trp Phe
1 5 10 15
Asp Pro
<210> 23
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 23
Ala Arg Ser Thr Gly Ser Gly Trp Thr His Asp Ala Phe Asp Ile
1 5 10 15
<210> 24
<211> 17
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 24
Val Arg Leu Asp Ser Asn Tyr Glu Asp Ser Gly Tyr Arg Ser Phe Asp
1 5 10 15
Arg
<210> 25
<211> 22
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 25
Val Thr Asp Arg Ser Phe Tyr Gly Ser Gly Thr Tyr Pro His Gln Phe
1 5 10 15
Tyr His Gly Leu Asp Val
20
<210> 26
<211> 14
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 26
Ala Ser Pro Leu Leu Trp Phe Gly Gly Tyr Tyr Met Asp Val
1 5 10
<210> 27
<211> 14
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 27
Ala Ser Pro Leu Leu Trp Phe Gly Gly Tyr Tyr Met Asp Val
1 5 10
<210> 28
<211> 26
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 28
Ala Arg Val Arg Glu Gly Ala Thr Asp Val Val Ala Val Pro Ala Ala
1 5 10 15
Thr Gly Tyr Tyr Ser Tyr Tyr Ile Asp Val
20 25
<210> 29
<211> 14
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 29
Ala Arg Val Pro Tyr Tyr Gly Asp Ser Thr Ile Phe Asp Tyr
1 5 10
<210> 30
<211> 11
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 30
Ala Arg Val Ser Gly Tyr Asp Ala Tyr Asp Tyr
1 5 10
<210> 31
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 31
Ala Arg Val Thr Ser Gly Asp Ser Phe Phe Tyr Tyr Tyr Met Asp Val
1 5 10 15
<210> 32
<211> 22
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 32
Ala Arg Gly Asp Tyr Asp Phe Trp Ser Gly Thr Ser Gly Lys Tyr Tyr
1 5 10 15
Ser Tyr Tyr Leu Asp Val
20
<210> 33
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 33
Ala Arg Val Thr Ser Gly Glu Ser Phe Phe Tyr Tyr Tyr Met Asp Val
1 5 10 15
<210> 34
<211> 17
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 34
Ala Arg Val Asp Leu Tyr Tyr Glu Asn Ser Gly Tyr Arg Ser Phe Asp
1 5 10 15
Ser
<210> 35
<211> 16
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 35
Ala Arg Thr Gly Tyr Ser Ser Gly Trp Tyr Glu Val Gly Leu Asp Tyr
1 5 10 15
<210> 36
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 36
Ala Arg Gln Ser Tyr Gly Thr Gly Ser Tyr Tyr Asp Phe Asp Ile
1 5 10 15
<210> 37
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 37
Ala Arg Gln Ser Tyr Gly Thr Gly Ser Tyr Tyr Asp Phe Asp Val
1 5 10 15
<210> 38
<211> 15
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 38
Ala Thr Asp Arg Pro Val Gly Val Val Ile Tyr Ala Leu Asp Phe
1 5 10 15
<210> 39
<211> 13
<212> PRT
<213> 人外周血(Human Peripheral Blood )
<400> 39
Val Lys Thr Gly Thr Thr Glu Asn Gln His Phe Asp Leu
1 5 10
Claims (4)
1.一种筛选特异性BCR/TCR的方法,其特征在于,包括以下步骤:
(1)分别提取N个免疫后的待测样本淋巴细胞中的mRNA;
(2)逆转录处理步骤(1)所得mRNA后,再对其BCR/TCR可变区基因进行多重PCR扩增,然后进行高通量测序,并确定BCR/TCR可变区的核酸及氨基酸序列;
(3)将测序得到的序列与未经抗原刺激样本的淋巴细胞BCR/TCR测序结果对比,获得相应的特异性免疫序列;
其中,包括所述将所有免疫后样本的BCR/TCR序列归总去重,再去除所有与未经抗原刺激样本的BCR/TCR测序结果序列集中所包含序列相重复的序列,建立特征性BCR/TCR序列集的步骤;
所述步骤(3)中特异性免疫序列的具体获得方式为:
(1)从特征性BCR/TCR序列集中去除只在<N×10%个免疫后样本中出现的序列;
(2)对特征性BCR/TCR序列集中所含有的共M种具有唯一性的BCR/TCR抗原决定簇3CDR3序列分别在N个免疫后样本测序结果中出现的次数进行统计,计为X1,1、X1,2、X1,3······X1,M,X2,1、X2,2、X2,3······X2,M,······,XN,1、XN,2、XN,3······XN,M;
(3)将特征性序列集中相同BCR/TCR CDR3序列在N个免疫后样本测序结果中出现次数求和,第j种CDR3序列出现次数之和为:
(4)将特征性序列集中所含有的共M种BCR/TCR CDR3序列按Yj值从高至低排序,排序前100个或前M×10%个序列即为与抗原相对应的特异性BCR/TCR CDR3序列。
2.根据权利要求1所述的筛选特异性BCR/TCR的方法,其特征在于,所述待测样本为包含淋巴细胞的外周血、脾脏组织、肿瘤组织或体外培养的淋巴细胞。
3.根据权利要求1或2任一项所述的筛选特异性BCR/TCR的方法,其特征在于,所述待测样本为人源样本或动物源样本。
4.一种用于筛选特异性BCR/TCR的系统,其特征在于,所述系统采用权利要求1-3任一项所述的方法对特异性BCR/TCR进行筛选。
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110273631.0A CN113122617B (zh) | 2021-03-15 | 2021-03-15 | 一种筛选特异bcr/tcr的方法及其系统 |
PCT/CN2022/080277 WO2022194032A1 (zh) | 2021-03-15 | 2022-03-11 | 一种筛选特异bcr/tcr的方法及其系统 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110273631.0A CN113122617B (zh) | 2021-03-15 | 2021-03-15 | 一种筛选特异bcr/tcr的方法及其系统 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113122617A CN113122617A (zh) | 2021-07-16 |
CN113122617B true CN113122617B (zh) | 2023-07-14 |
Family
ID=76773362
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110273631.0A Active CN113122617B (zh) | 2021-03-15 | 2021-03-15 | 一种筛选特异bcr/tcr的方法及其系统 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN113122617B (zh) |
WO (1) | WO2022194032A1 (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113122617B (zh) * | 2021-03-15 | 2023-07-14 | 成都益安博生物技术有限公司 | 一种筛选特异bcr/tcr的方法及其系统 |
CN114349847B (zh) * | 2022-02-08 | 2023-07-21 | 深圳市因诺转化医学研究院 | 靶向新型冠状病毒rna依赖性rna聚合酶的特异性tcr |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH09127119A (ja) * | 1995-11-01 | 1997-05-16 | L T T Kenkyusho:Kk | C型慢性肝炎の治療効果判定方法 |
US8628927B2 (en) * | 2008-11-07 | 2014-01-14 | Sequenta, Inc. | Monitoring health and disease status using clonotype profiles |
US8748103B2 (en) * | 2008-11-07 | 2014-06-10 | Sequenta, Inc. | Monitoring health and disease status using clonotype profiles |
WO2014121272A2 (en) * | 2013-02-04 | 2014-08-07 | Quake Stephen R | Measurement and comparison of immune diversity by high-throughput sequencing |
US20160186260A1 (en) * | 2013-07-26 | 2016-06-30 | Sequenta, Llc | Cancer vaccination with antigen evolution |
US11203783B2 (en) * | 2013-11-21 | 2021-12-21 | Repertoire Genesis Incorporation | T cell receptor and B cell receptor repertoire analysis system, and use of same in treatment and diagnosis |
CN106156536B (zh) * | 2015-04-15 | 2018-11-13 | 深圳华大基因科技有限公司 | 对样本免疫组库测序数据进行处理的方法和系统 |
WO2017193097A1 (en) * | 2016-05-06 | 2017-11-09 | Girihlet Inc. | Methods and compositions for determining specifc tcr and bcr chain pairings |
US10822662B2 (en) * | 2017-03-06 | 2020-11-03 | Karkinos Precision Oncology LLC | Diagnostic methods for identifying T-cell lymphoma and leukemia by high-throughput TCR-β sequencing |
US20200199650A1 (en) * | 2017-05-18 | 2020-06-25 | Geneplus-Beijing | Analysis system for peripheral blood-based non-invasive detection of lesion immune repertoire diversity and uses of system |
CN110246539B (zh) * | 2019-04-15 | 2020-04-28 | 成都益安博生物技术有限公司 | 一种免疫力水平评估的方法及装置 |
CN110257476A (zh) * | 2019-05-31 | 2019-09-20 | 南方医科大学南方医院 | 一种甄别样品间交叉反应的免疫组库高通量测序文库的构建方法 |
CN113122617B (zh) * | 2021-03-15 | 2023-07-14 | 成都益安博生物技术有限公司 | 一种筛选特异bcr/tcr的方法及其系统 |
-
2021
- 2021-03-15 CN CN202110273631.0A patent/CN113122617B/zh active Active
-
2022
- 2022-03-11 WO PCT/CN2022/080277 patent/WO2022194032A1/zh active Application Filing
Also Published As
Publication number | Publication date |
---|---|
WO2022194032A1 (zh) | 2022-09-22 |
CN113122617A (zh) | 2021-07-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Sutton et al. | Atypical B cells are part of an alternative lineage of B cells that participates in responses to vaccination and infection in humans | |
CN115304671B (zh) | 新冠病毒rbd特异性单克隆抗体及其应用 | |
CN113122617B (zh) | 一种筛选特异bcr/tcr的方法及其系统 | |
AU776464B2 (en) | Monoclonal antibodies, antigens and diagnosis and therapy of malignant diseases | |
CN114920832B (zh) | 新冠病毒rbd特异性单克隆抗体和应用 | |
CN114920833B (zh) | 新冠病毒rbd特异性单克隆抗体和应用 | |
KR20130135028A (ko) | 동물로부터 단일클론 항체의 고속 분리 | |
CA2471570A1 (en) | Focussed antibody technology | |
CN115925902A (zh) | 新型冠状病毒rbd特异性单克隆抗体和应用 | |
CN111040031B (zh) | 一种抗非洲猪瘟病毒的ScFv抗体及其制备方法 | |
CN114989293A (zh) | 新冠病毒rbd特异性单克隆抗体和应用 | |
JP2002532066A (ja) | 鳥類におけるポリヌクレオチドワクチンを用いる抗体の産生 | |
US20230091895A1 (en) | Nanobody for pcsk9 and application thereof | |
CN112661841A (zh) | 一种中和新冠病毒新表位的全人单克隆抗体17-2及其应用 | |
CN114349855A (zh) | 新型冠状病毒Delta突变株特异性抗体及其应用 | |
CN111647070B (zh) | T细胞受体或其抗原结合片段及应用 | |
CN110655572B (zh) | 一种抗丝状病毒gp蛋白的单克隆抗体及其应用 | |
CN114409791A (zh) | 一种全人源抗人红细胞RhD全分子IgG及其制备方法和应用 | |
CN114729403A (zh) | 用于鉴定和验证共有候选抗原和共有抗原特异性t淋巴细胞对的方法和系统 | |
CN108822189A (zh) | 一种靶向结合淋巴癌细胞系的特异性多肽及其应用 | |
CN114957455B (zh) | 一种新型冠状病毒单克隆抗体及其应用 | |
CN111647069B (zh) | 一种改进的tcr及其应用 | |
CN108484770A (zh) | 重组大鼠抗小鼠cd4单克隆抗体,制备方法和应用 | |
CN116199779B (zh) | 一种抗lilrb4单克隆抗体、其抗原结合片段及其应用 | |
CN116836273B (zh) | 抗血清淀粉样蛋白a抗体、检测血清淀粉样蛋白a的试剂和试剂盒 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |