WO2006137932A2 - Homogeneous analyte detection - Google Patents
Homogeneous analyte detection Download PDFInfo
- Publication number
- WO2006137932A2 WO2006137932A2 PCT/US2005/040133 US2005040133W WO2006137932A2 WO 2006137932 A2 WO2006137932 A2 WO 2006137932A2 US 2005040133 W US2005040133 W US 2005040133W WO 2006137932 A2 WO2006137932 A2 WO 2006137932A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- nucleic acid
- binding
- duplex
- binding pair
- dna
- Prior art date
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 41
- 239000012491 analyte Substances 0.000 title claims description 81
- 238000009739 binding Methods 0.000 claims abstract description 252
- 230000027455 binding Effects 0.000 claims abstract description 251
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 202
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 197
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 197
- 238000000034 method Methods 0.000 claims abstract description 123
- 210000004027 cell Anatomy 0.000 claims description 48
- 108091007433 antigens Proteins 0.000 claims description 36
- 102000036639 antigens Human genes 0.000 claims description 36
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 35
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 34
- 230000003321 amplification Effects 0.000 claims description 33
- 239000000427 antigen Substances 0.000 claims description 32
- 108010072866 Prostate-Specific Antigen Proteins 0.000 claims description 29
- 102100038358 Prostate-specific antigen Human genes 0.000 claims description 29
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 19
- 230000000295 complement effect Effects 0.000 claims description 18
- 239000003446 ligand Substances 0.000 claims description 15
- 239000000243 solution Substances 0.000 claims description 15
- 210000001519 tissue Anatomy 0.000 claims description 13
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 claims description 11
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 claims description 11
- 239000002773 nucleotide Substances 0.000 claims description 10
- 125000003729 nucleotide group Chemical group 0.000 claims description 9
- 108020004414 DNA Proteins 0.000 claims description 8
- 230000008018 melting Effects 0.000 claims description 8
- 238000002844 melting Methods 0.000 claims description 8
- 150000001875 compounds Chemical class 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 7
- 230000001580 bacterial effect Effects 0.000 claims description 6
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 claims description 6
- 229960005542 ethidium bromide Drugs 0.000 claims description 6
- 238000010438 heat treatment Methods 0.000 claims description 6
- 125000006850 spacer group Chemical group 0.000 claims description 5
- 238000010186 staining Methods 0.000 claims description 5
- 102000006382 Ribonucleases Human genes 0.000 claims description 4
- 108010083644 Ribonucleases Proteins 0.000 claims description 4
- 210000004102 animal cell Anatomy 0.000 claims description 4
- 238000007865 diluting Methods 0.000 claims description 4
- 208000002109 Argyria Diseases 0.000 claims description 3
- 238000002105 Southern blotting Methods 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 238000000376 autoradiography Methods 0.000 claims description 3
- 210000004369 blood Anatomy 0.000 claims description 3
- 239000008280 blood Substances 0.000 claims description 3
- 230000002538 fungal effect Effects 0.000 claims description 3
- 210000005260 human cell Anatomy 0.000 claims description 3
- 210000002966 serum Anatomy 0.000 claims description 3
- 230000003247 decreasing effect Effects 0.000 claims description 2
- 230000002285 radioactive effect Effects 0.000 claims description 2
- 238000003752 polymerase chain reaction Methods 0.000 claims 2
- 239000000203 mixture Substances 0.000 abstract description 22
- 238000011896 sensitive detection Methods 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 description 47
- 239000013615 primer Substances 0.000 description 43
- 235000018102 proteins Nutrition 0.000 description 41
- 102000004169 proteins and genes Human genes 0.000 description 41
- 238000003556 assay Methods 0.000 description 39
- 239000003153 chemical reaction reagent Substances 0.000 description 36
- 108091034117 Oligonucleotide Proteins 0.000 description 29
- 102000005962 receptors Human genes 0.000 description 24
- 108020003175 receptors Proteins 0.000 description 24
- 108090000765 processed proteins & peptides Proteins 0.000 description 20
- 239000000306 component Substances 0.000 description 18
- 238000006243 chemical reaction Methods 0.000 description 17
- 102000004196 processed proteins & peptides Human genes 0.000 description 17
- 230000035945 sensitivity Effects 0.000 description 16
- 241000700605 Viruses Species 0.000 description 15
- 230000015572 biosynthetic process Effects 0.000 description 15
- 238000005755 formation reaction Methods 0.000 description 15
- 239000000523 sample Substances 0.000 description 15
- 238000009396 hybridization Methods 0.000 description 14
- 239000003550 marker Substances 0.000 description 13
- 229920001184 polypeptide Polymers 0.000 description 13
- 238000003753 real-time PCR Methods 0.000 description 12
- 239000000126 substance Substances 0.000 description 12
- 230000021615 conjugation Effects 0.000 description 11
- 239000007787 solid Substances 0.000 description 11
- 238000003018 immunoassay Methods 0.000 description 10
- -1 poly(amino acids) Polymers 0.000 description 10
- 241001465754 Metazoa Species 0.000 description 9
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 8
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 239000012634 fragment Substances 0.000 description 8
- 230000006870 function Effects 0.000 description 8
- 239000007790 solid phase Substances 0.000 description 8
- 206010028980 Neoplasm Diseases 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 7
- 150000001413 amino acids Chemical class 0.000 description 7
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 7
- 230000000670 limiting effect Effects 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 230000009871 nonspecific binding Effects 0.000 description 7
- 108091033319 polynucleotide Proteins 0.000 description 7
- 102000040430 polynucleotide Human genes 0.000 description 7
- 239000002157 polynucleotide Substances 0.000 description 7
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 7
- 241000588724 Escherichia coli Species 0.000 description 6
- 108060003951 Immunoglobulin Proteins 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 108010090804 Streptavidin Proteins 0.000 description 6
- 239000007983 Tris buffer Substances 0.000 description 6
- 102000018358 immunoglobulin Human genes 0.000 description 6
- 244000005700 microbiome Species 0.000 description 6
- 210000002381 plasma Anatomy 0.000 description 6
- 230000009870 specific binding Effects 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 239000011324 bead Substances 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- 239000008364 bulk solution Substances 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 230000006862 enzymatic digestion Effects 0.000 description 5
- 229940088598 enzyme Drugs 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 229940088597 hormone Drugs 0.000 description 5
- 239000005556 hormone Substances 0.000 description 5
- 230000003993 interaction Effects 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- 150000008300 phosphoramidites Chemical class 0.000 description 5
- 239000004033 plastic Substances 0.000 description 5
- 229920003023 plastic Polymers 0.000 description 5
- TYFQFVWCELRYAO-UHFFFAOYSA-L suberate(2-) Chemical compound [O-]C(=O)CCCCCCC([O-])=O TYFQFVWCELRYAO-UHFFFAOYSA-L 0.000 description 5
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 5
- ASJSAQIRZKANQN-CRCLSJGQSA-N 2-deoxy-D-ribose Chemical compound OC[C@@H](O)[C@@H](O)CC=O ASJSAQIRZKANQN-CRCLSJGQSA-N 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- 102000053602 DNA Human genes 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 4
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 4
- 229960002685 biotin Drugs 0.000 description 4
- 235000020958 biotin Nutrition 0.000 description 4
- 239000011616 biotin Substances 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 230000009918 complex formation Effects 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 210000004408 hybridoma Anatomy 0.000 description 4
- 229940027941 immunoglobulin g Drugs 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 4
- 238000002372 labelling Methods 0.000 description 4
- 230000005298 paramagnetic effect Effects 0.000 description 4
- 238000006116 polymerization reaction Methods 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 241000304886 Bacilli Species 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 102000006395 Globulins Human genes 0.000 description 3
- 108010044091 Globulins Proteins 0.000 description 3
- 108010024636 Glutathione Proteins 0.000 description 3
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 3
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 3
- HDFGOPSGAURCEO-UHFFFAOYSA-N N-ethylmaleimide Chemical compound CCN1C(=O)C=CC1=O HDFGOPSGAURCEO-UHFFFAOYSA-N 0.000 description 3
- 239000004677 Nylon Substances 0.000 description 3
- 241000607768 Shigella Species 0.000 description 3
- 108010006785 Taq Polymerase Proteins 0.000 description 3
- 102000004987 Troponin T Human genes 0.000 description 3
- 108090001108 Troponin T Proteins 0.000 description 3
- 230000000890 antigenic effect Effects 0.000 description 3
- 239000013060 biological fluid Substances 0.000 description 3
- 230000001413 cellular effect Effects 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
- 239000002299 complementary DNA Substances 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 229960003180 glutathione Drugs 0.000 description 3
- 229940099472 immunoglobulin a Drugs 0.000 description 3
- 238000010348 incorporation Methods 0.000 description 3
- 230000000977 initiatory effect Effects 0.000 description 3
- 239000004816 latex Substances 0.000 description 3
- 229920000126 latex Polymers 0.000 description 3
- 230000005291 magnetic effect Effects 0.000 description 3
- 239000011859 microparticle Substances 0.000 description 3
- 239000000178 monomer Substances 0.000 description 3
- 229920001778 nylon Polymers 0.000 description 3
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 3
- 239000002987 primer (paints) Substances 0.000 description 3
- 239000011541 reaction mixture Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000000108 ultra-filtration Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- HPZMWTNATZPBIH-UHFFFAOYSA-N 1-methyladenine Chemical compound CN1C=NC2=NC=NC2=C1N HPZMWTNATZPBIH-UHFFFAOYSA-N 0.000 description 2
- RFLVMTUMFYRZCB-UHFFFAOYSA-N 1-methylguanine Chemical compound O=C1N(C)C(N)=NC2=C1N=CN2 RFLVMTUMFYRZCB-UHFFFAOYSA-N 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- 108090001008 Avidin Proteins 0.000 description 2
- 108060001064 Calcitonin Proteins 0.000 description 2
- 102000055006 Calcitonin Human genes 0.000 description 2
- 241000606161 Chlamydia Species 0.000 description 2
- 108020004635 Complementary DNA Proteins 0.000 description 2
- 102400000739 Corticotropin Human genes 0.000 description 2
- 101800000414 Corticotropin Proteins 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 2
- 108010017826 DNA Polymerase I Proteins 0.000 description 2
- 102000004594 DNA Polymerase I Human genes 0.000 description 2
- 239000003155 DNA primer Substances 0.000 description 2
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 2
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 102000003951 Erythropoietin Human genes 0.000 description 2
- 108090000394 Erythropoietin Proteins 0.000 description 2
- 108010014173 Factor X Proteins 0.000 description 2
- 108010049003 Fibrinogen Proteins 0.000 description 2
- 102000008946 Fibrinogen Human genes 0.000 description 2
- 108010051696 Growth Hormone Proteins 0.000 description 2
- 208000007514 Herpes zoster Diseases 0.000 description 2
- 108091027305 Heteroduplex Proteins 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 2
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 2
- 102000004877 Insulin Human genes 0.000 description 2
- 108090001061 Insulin Proteins 0.000 description 2
- 102000003746 Insulin Receptor Human genes 0.000 description 2
- 108010001127 Insulin Receptor Proteins 0.000 description 2
- 102100034343 Integrase Human genes 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 241000144128 Lichtheimia corymbifera Species 0.000 description 2
- 102000009151 Luteinizing Hormone Human genes 0.000 description 2
- 108010073521 Luteinizing Hormone Proteins 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 108010007013 Melanocyte-Stimulating Hormones Proteins 0.000 description 2
- 102100030856 Myoglobin Human genes 0.000 description 2
- 108010062374 Myoglobin Proteins 0.000 description 2
- HYVABZIGRDEKCD-UHFFFAOYSA-N N(6)-dimethylallyladenine Chemical compound CC(C)=CCNC1=NC=NC2=C1N=CN2 HYVABZIGRDEKCD-UHFFFAOYSA-N 0.000 description 2
- GHAZCVNUKKZTLG-UHFFFAOYSA-N N-ethyl-succinimide Natural products CCN1C(=O)CCC1=O GHAZCVNUKKZTLG-UHFFFAOYSA-N 0.000 description 2
- 239000000020 Nitrocellulose Substances 0.000 description 2
- 108020004711 Nucleic Acid Probes Proteins 0.000 description 2
- 208000002606 Paramyxoviridae Infections Diseases 0.000 description 2
- 239000004793 Polystyrene Substances 0.000 description 2
- 108091028664 Ribonucleotide Proteins 0.000 description 2
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 2
- 241000606701 Rickettsia Species 0.000 description 2
- 241000606697 Rickettsia prowazekii Species 0.000 description 2
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 2
- 102100038803 Somatotropin Human genes 0.000 description 2
- 108010000499 Thromboplastin Proteins 0.000 description 2
- 102000002262 Thromboplastin Human genes 0.000 description 2
- 206010053614 Type III immune complex mediated reaction Diseases 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 230000005875 antibody response Effects 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000002820 assay format Methods 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 239000003114 blood coagulation factor Substances 0.000 description 2
- 238000010804 cDNA synthesis Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000005465 channeling Effects 0.000 description 2
- IDLFZVILOHSSID-OVLDLUHVSA-N corticotropin Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)NC(=O)[C@@H](N)CO)C1=CC=C(O)C=C1 IDLFZVILOHSSID-OVLDLUHVSA-N 0.000 description 2
- 229960000258 corticotropin Drugs 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
- 239000005547 deoxyribonucleotide Substances 0.000 description 2
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 210000003743 erythrocyte Anatomy 0.000 description 2
- 229940105423 erythropoietin Drugs 0.000 description 2
- 229940012952 fibrinogen Drugs 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000002866 fluorescence resonance energy transfer Methods 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 238000001641 gel filtration chromatography Methods 0.000 description 2
- 230000013595 glycosylation Effects 0.000 description 2
- 238000006206 glycosylation reaction Methods 0.000 description 2
- 230000016178 immune complex formation Effects 0.000 description 2
- 229940072221 immunoglobulins Drugs 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 229940125396 insulin Drugs 0.000 description 2
- 238000009830 intercalation Methods 0.000 description 2
- 208000032839 leukemia Diseases 0.000 description 2
- 125000005647 linker group Chemical group 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 210000003470 mitochondria Anatomy 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 208000010125 myocardial infarction Diseases 0.000 description 2
- 239000002858 neurotransmitter agent Substances 0.000 description 2
- 229920001220 nitrocellulos Polymers 0.000 description 2
- 239000002853 nucleic acid probe Substances 0.000 description 2
- 238000001668 nucleic acid synthesis Methods 0.000 description 2
- 210000003463 organelle Anatomy 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 239000000813 peptide hormone Substances 0.000 description 2
- RXNXLAHQOVLMIE-UHFFFAOYSA-N phenyl 10-methylacridin-10-ium-9-carboxylate Chemical compound C12=CC=CC=C2[N+](C)=C2C=CC=CC2=C1C(=O)OC1=CC=CC=C1 RXNXLAHQOVLMIE-UHFFFAOYSA-N 0.000 description 2
- 230000026731 phosphorylation Effects 0.000 description 2
- 238000006366 phosphorylation reaction Methods 0.000 description 2
- 229920002223 polystyrene Polymers 0.000 description 2
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 210000002307 prostate Anatomy 0.000 description 2
- 208000017497 prostate disease Diseases 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 239000003488 releasing hormone Substances 0.000 description 2
- 239000002336 ribonucleotide Substances 0.000 description 2
- 125000002652 ribonucleotide group Chemical group 0.000 description 2
- 150000003290 ribose derivatives Chemical class 0.000 description 2
- 229940046939 rickettsia prowazekii Drugs 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000001488 sodium phosphate Substances 0.000 description 2
- 229910000162 sodium phosphate Inorganic materials 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- 239000003053 toxin Substances 0.000 description 2
- 231100000765 toxin Toxicity 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 238000012384 transportation and delivery Methods 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 239000001226 triphosphate Substances 0.000 description 2
- 235000011178 triphosphate Nutrition 0.000 description 2
- 239000003656 tris buffered saline Substances 0.000 description 2
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 2
- 210000002700 urine Anatomy 0.000 description 2
- 238000010792 warming Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- SFVVQRJOGUKCEG-OPQSFPLASA-N β-MSH Chemical compound C1C[C@@H](O)[C@H]2C(COC(=O)[C@@](O)([C@@H](C)O)C(C)C)=CCN21 SFVVQRJOGUKCEG-OPQSFPLASA-N 0.000 description 2
- PGOHTUIFYSHAQG-LJSDBVFPSA-N (2S)-6-amino-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-1-[(2S,3R)-2-[[(2S)-2-[[(2S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-1-[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-4-methylsulfanylbutanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-5-carbamimidamidopentanoyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]-3-methylbutanoyl]amino]-4-methylpentanoyl]amino]-4-methylpentanoyl]amino]acetyl]amino]-3-hydroxypropanoyl]amino]-4-methylpentanoyl]amino]-3-sulfanylpropanoyl]amino]-4-methylsulfanylbutanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-hydroxybutanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-hydroxypropanoyl]amino]-3-hydroxypropanoyl]amino]-3-(1H-imidazol-5-yl)propanoyl]amino]-4-methylpentanoyl]amino]-3-hydroxybutanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-oxopentanoyl]amino]-3-hydroxybutanoyl]amino]-3-hydroxypropanoyl]amino]-3-carboxypropanoyl]amino]-3-hydroxypropanoyl]amino]-5-oxopentanoyl]amino]-5-oxopentanoyl]amino]-3-phenylpropanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-methylbutanoyl]amino]-4-methylpentanoyl]amino]-4-oxobutanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-4-carboxybutanoyl]amino]-5-oxopentanoyl]amino]hexanoic acid Chemical compound CSCC[C@H](N)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(O)=O PGOHTUIFYSHAQG-LJSDBVFPSA-N 0.000 description 1
- CUKWUWBLQQDQAC-VEQWQPCFSA-N (3s)-3-amino-4-[[(2s)-1-[[(2s)-1-[[(2s)-1-[[(2s,3s)-1-[[(2s)-1-[(2s)-2-[[(1s)-1-carboxyethyl]carbamoyl]pyrrolidin-1-yl]-3-(1h-imidazol-5-yl)-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-3-methyl-1-ox Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C1=CC=C(O)C=C1 CUKWUWBLQQDQAC-VEQWQPCFSA-N 0.000 description 1
- SATCOUWSAZBIJO-UHFFFAOYSA-N 1-methyladenine Natural products N=C1N(C)C=NC2=C1NC=N2 SATCOUWSAZBIJO-UHFFFAOYSA-N 0.000 description 1
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 description 1
- WJNGQIYEQLPJMN-IOSLPCCCSA-N 1-methylinosine Chemical compound C1=NC=2C(=O)N(C)C=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O WJNGQIYEQLPJMN-IOSLPCCCSA-N 0.000 description 1
- GZCWLCBFPRFLKL-UHFFFAOYSA-N 1-prop-2-ynoxypropan-2-ol Chemical compound CC(O)COCC#C GZCWLCBFPRFLKL-UHFFFAOYSA-N 0.000 description 1
- SVBOROZXXYRWJL-UHFFFAOYSA-N 2-[(4-oxo-2-sulfanylidene-1h-pyrimidin-5-yl)methylamino]acetic acid Chemical compound OC(=O)CNCC1=CNC(=S)NC1=O SVBOROZXXYRWJL-UHFFFAOYSA-N 0.000 description 1
- LLWPKTDSDUQBFY-UHFFFAOYSA-N 2-[6-(aminomethyl)-2,4-dioxo-1H-pyrimidin-5-yl]acetic acid Chemical compound C(=O)(O)CC=1C(NC(NC=1CN)=O)=O LLWPKTDSDUQBFY-UHFFFAOYSA-N 0.000 description 1
- KISWVXRQTGLFGD-UHFFFAOYSA-N 2-[[2-[[6-amino-2-[[2-[[2-[[5-amino-2-[[2-[[1-[2-[[6-amino-2-[(2,5-diamino-5-oxopentanoyl)amino]hexanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxypropanoyl]amino]-5-oxopentanoyl]amino]-5-(diaminomethylideneamino)p Chemical compound C1CCN(C(=O)C(CCCN=C(N)N)NC(=O)C(CCCCN)NC(=O)C(N)CCC(N)=O)C1C(=O)NC(CO)C(=O)NC(CCC(N)=O)C(=O)NC(CCCN=C(N)N)C(=O)NC(CO)C(=O)NC(CCCCN)C(=O)NC(C(=O)NC(CC(C)C)C(O)=O)CC1=CC=C(O)C=C1 KISWVXRQTGLFGD-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- XMSMHKMPBNTBOD-UHFFFAOYSA-N 2-dimethylamino-6-hydroxypurine Chemical compound N1C(N(C)C)=NC(=O)C2=C1N=CN2 XMSMHKMPBNTBOD-UHFFFAOYSA-N 0.000 description 1
- SMADWRYCYBUIKH-UHFFFAOYSA-N 2-methyl-7h-purin-6-amine Chemical compound CC1=NC(N)=C2NC=NC2=N1 SMADWRYCYBUIKH-UHFFFAOYSA-N 0.000 description 1
- KOLPWZCZXAMXKS-UHFFFAOYSA-N 3-methylcytosine Chemical compound CN1C(N)=CC=NC1=O KOLPWZCZXAMXKS-UHFFFAOYSA-N 0.000 description 1
- GJAKJCICANKRFD-UHFFFAOYSA-N 4-acetyl-4-amino-1,3-dihydropyrimidin-2-one Chemical compound CC(=O)C1(N)NC(=O)NC=C1 GJAKJCICANKRFD-UHFFFAOYSA-N 0.000 description 1
- LQLQRFGHAALLLE-UHFFFAOYSA-N 5-bromouracil Chemical compound BrC1=CNC(=O)NC1=O LQLQRFGHAALLLE-UHFFFAOYSA-N 0.000 description 1
- LRSASMSXMSNRBT-UHFFFAOYSA-N 5-methylcytosine Chemical compound CC1=CNC(=O)N=C1N LRSASMSXMSNRBT-UHFFFAOYSA-N 0.000 description 1
- MSSXOMSJDRHRMC-UHFFFAOYSA-N 9H-purine-2,6-diamine Chemical compound NC1=NC(N)=C2NC=NC2=N1 MSSXOMSJDRHRMC-UHFFFAOYSA-N 0.000 description 1
- 102000013563 Acid Phosphatase Human genes 0.000 description 1
- 108010051457 Acid Phosphatase Proteins 0.000 description 1
- 241000186046 Actinomyces Species 0.000 description 1
- 241000186042 Actinomyces bovis Species 0.000 description 1
- 241000186045 Actinomyces naeslundii Species 0.000 description 1
- 239000000275 Adrenocorticotropic Hormone Substances 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 241000588813 Alcaligenes faecalis Species 0.000 description 1
- 102400000344 Angiotensin-1 Human genes 0.000 description 1
- 101800000734 Angiotensin-1 Proteins 0.000 description 1
- 102400000345 Angiotensin-2 Human genes 0.000 description 1
- 101800000733 Angiotensin-2 Proteins 0.000 description 1
- 206010003445 Ascites Diseases 0.000 description 1
- 241001225321 Aspergillus fumigatus Species 0.000 description 1
- 241000351920 Aspergillus nidulans Species 0.000 description 1
- 241000711404 Avian avulavirus 1 Species 0.000 description 1
- 238000000035 BCA protein assay Methods 0.000 description 1
- 241000193738 Bacillus anthracis Species 0.000 description 1
- 241000193755 Bacillus cereus Species 0.000 description 1
- 241000194107 Bacillus megaterium Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000335423 Blastomyces Species 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 241000180135 Borrelia recurrentis Species 0.000 description 1
- 208000003508 Botulism Diseases 0.000 description 1
- 102400000967 Bradykinin Human genes 0.000 description 1
- 101800004538 Bradykinin Proteins 0.000 description 1
- 241000589567 Brucella abortus Species 0.000 description 1
- 241001148106 Brucella melitensis Species 0.000 description 1
- 241001148111 Brucella suis Species 0.000 description 1
- 108010074051 C-Reactive Protein Proteins 0.000 description 1
- 102100032752 C-reactive protein Human genes 0.000 description 1
- 241001493160 California encephalitis virus Species 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- 241000712083 Canine morbillivirus Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 102000053642 Catalytic RNA Human genes 0.000 description 1
- 108090000994 Catalytic RNA Proteins 0.000 description 1
- 102000000844 Cell Surface Receptors Human genes 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 108010075016 Ceruloplasmin Proteins 0.000 description 1
- 102100023321 Ceruloplasmin Human genes 0.000 description 1
- 201000006082 Chickenpox Diseases 0.000 description 1
- 108091092236 Chimeric RNA Proteins 0.000 description 1
- 102000003914 Cholinesterases Human genes 0.000 description 1
- 108090000322 Cholinesterases Proteins 0.000 description 1
- 102000011022 Chorionic Gonadotropin Human genes 0.000 description 1
- 108010062540 Chorionic Gonadotropin Proteins 0.000 description 1
- 108020004638 Circular DNA Proteins 0.000 description 1
- 241001668502 Cladophialophora carrionii Species 0.000 description 1
- 241000193155 Clostridium botulinum Species 0.000 description 1
- 241000186581 Clostridium novyi Species 0.000 description 1
- 241000193468 Clostridium perfringens Species 0.000 description 1
- 241000193466 Clostridium septicum Species 0.000 description 1
- 241000193470 Clostridium sporogenes Species 0.000 description 1
- 241000186528 Clostridium tertium Species 0.000 description 1
- 241000193449 Clostridium tetani Species 0.000 description 1
- 102100022641 Coagulation factor IX Human genes 0.000 description 1
- 102100023804 Coagulation factor VII Human genes 0.000 description 1
- 102100029117 Coagulation factor X Human genes 0.000 description 1
- 241000223205 Coccidioides immitis Species 0.000 description 1
- 241000204955 Colorado tick fever virus Species 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 241000186216 Corynebacterium Species 0.000 description 1
- 241000709687 Coxsackievirus Species 0.000 description 1
- 241000221204 Cryptococcus neoformans Species 0.000 description 1
- 241000701022 Cytomegalovirus Species 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 241000725619 Dengue virus Species 0.000 description 1
- 241000710945 Eastern equine encephalitis virus Species 0.000 description 1
- 241001466953 Echovirus Species 0.000 description 1
- 241000709661 Enterovirus Species 0.000 description 1
- 241000991587 Enterovirus C Species 0.000 description 1
- 101000867232 Escherichia coli Heat-stable enterotoxin II Proteins 0.000 description 1
- 241001646719 Escherichia coli O157:H7 Species 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- 241000223664 Exophiala jeanselmei Species 0.000 description 1
- 108010076282 Factor IX Proteins 0.000 description 1
- 108010014172 Factor V Proteins 0.000 description 1
- 108010023321 Factor VII Proteins 0.000 description 1
- 108010071289 Factor XIII Proteins 0.000 description 1
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 1
- 102000012673 Follicle Stimulating Hormone Human genes 0.000 description 1
- 108010079345 Follicle Stimulating Hormone Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 102400000921 Gastrin Human genes 0.000 description 1
- 108010052343 Gastrins Proteins 0.000 description 1
- 102400000321 Glucagon Human genes 0.000 description 1
- 108060003199 Glucagon Proteins 0.000 description 1
- 108010086677 Gonadotropins Proteins 0.000 description 1
- 102000006771 Gonadotropins Human genes 0.000 description 1
- QXZGBUJJYSLZLT-UHFFFAOYSA-N H-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-OH Natural products NC(N)=NCCCC(N)C(=O)N1CCCC1C(=O)N1C(C(=O)NCC(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CO)C(=O)N2C(CCC2)C(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CCCN=C(N)N)C(O)=O)CCC1 QXZGBUJJYSLZLT-UHFFFAOYSA-N 0.000 description 1
- 102100025255 Haptoglobin Human genes 0.000 description 1
- 108050005077 Haptoglobin Proteins 0.000 description 1
- 241000193159 Hathewaya histolytica Species 0.000 description 1
- 108010026027 Hemopexin Proteins 0.000 description 1
- 102000013271 Hemopexin Human genes 0.000 description 1
- 241000711549 Hepacivirus C Species 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 241000709721 Hepatovirus A Species 0.000 description 1
- 208000009889 Herpes Simplex Diseases 0.000 description 1
- 102000006947 Histones Human genes 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 101000837639 Homo sapiens Thyroxine-binding globulin Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 241000701806 Human papillomavirus Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 102000009786 Immunoglobulin Constant Regions Human genes 0.000 description 1
- 108010009817 Immunoglobulin Constant Regions Proteins 0.000 description 1
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 1
- 229930010555 Inosine Natural products 0.000 description 1
- 101710203526 Integrase Proteins 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 241001221585 Keratinomyces Species 0.000 description 1
- 241000588915 Klebsiella aerogenes Species 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- 102000007330 LDL Lipoproteins Human genes 0.000 description 1
- 108010007622 LDL Lipoproteins Proteins 0.000 description 1
- 241000589902 Leptospira Species 0.000 description 1
- 241001550390 Leptospira interrogans serovar Canicola Species 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241001444203 Madurella mycetomatis Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241000712079 Measles morbillivirus Species 0.000 description 1
- 239000000637 Melanocyte-Stimulating Hormone Substances 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 241001480037 Microsporum Species 0.000 description 1
- 241000893980 Microsporum canis Species 0.000 description 1
- 241001430197 Mollicutes Species 0.000 description 1
- 241000588772 Morganella morganii Species 0.000 description 1
- 108010093825 Mucoproteins Proteins 0.000 description 1
- 102000001621 Mucoproteins Human genes 0.000 description 1
- 241000711386 Mumps virus Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000186367 Mycobacterium avium Species 0.000 description 1
- 241000187482 Mycobacterium avium subsp. paratuberculosis Species 0.000 description 1
- 241000186366 Mycobacterium bovis Species 0.000 description 1
- 241000186362 Mycobacterium leprae Species 0.000 description 1
- 241000187479 Mycobacterium tuberculosis Species 0.000 description 1
- 241000202934 Mycoplasma pneumoniae Species 0.000 description 1
- 102000047918 Myelin Basic Human genes 0.000 description 1
- 101710107068 Myelin basic protein Proteins 0.000 description 1
- SGSSKEDGVONRGC-UHFFFAOYSA-N N(2)-methylguanine Chemical compound O=C1NC(NC)=NC2=C1N=CN2 SGSSKEDGVONRGC-UHFFFAOYSA-N 0.000 description 1
- 241000893976 Nannizzia gypsea Species 0.000 description 1
- 241000588653 Neisseria Species 0.000 description 1
- 241000588652 Neisseria gonorrhoeae Species 0.000 description 1
- 241000588650 Neisseria meningitidis Species 0.000 description 1
- 241000187678 Nocardia asteroides Species 0.000 description 1
- 241001503696 Nocardia brasiliensis Species 0.000 description 1
- 102000007399 Nuclear hormone receptor Human genes 0.000 description 1
- 241000606693 Orientia tsutsugamushi Species 0.000 description 1
- 241000702244 Orthoreovirus Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 102400000050 Oxytocin Human genes 0.000 description 1
- 101800000989 Oxytocin Proteins 0.000 description 1
- XNOPRXBHLZRZKH-UHFFFAOYSA-N Oxytocin Natural products N1C(=O)C(N)CSSCC(C(=O)N2C(CCC2)C(=O)NC(CC(C)C)C(=O)NCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(CCC(N)=O)NC(=O)C(C(C)CC)NC(=O)C1CC1=CC=C(O)C=C1 XNOPRXBHLZRZKH-UHFFFAOYSA-N 0.000 description 1
- 239000012807 PCR reagent Substances 0.000 description 1
- 241000193157 Paraclostridium bifermentans Species 0.000 description 1
- 241000526686 Paracoccidioides brasiliensis Species 0.000 description 1
- 102000003982 Parathyroid hormone Human genes 0.000 description 1
- 108090000445 Parathyroid hormone Proteins 0.000 description 1
- 206010033976 Paravaccinia Diseases 0.000 description 1
- 241000606860 Pasteurella Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 201000005702 Pertussis Diseases 0.000 description 1
- 241001531356 Phialophora verrucosa Species 0.000 description 1
- 108010089430 Phosphoproteins Proteins 0.000 description 1
- 102000007982 Phosphoproteins Human genes 0.000 description 1
- 241000709664 Picornaviridae Species 0.000 description 1
- 102000004576 Placental Lactogen Human genes 0.000 description 1
- 108010003044 Placental Lactogen Proteins 0.000 description 1
- 239000000381 Placental Lactogen Substances 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 102100038124 Plasminogen Human genes 0.000 description 1
- 108010051456 Plasminogen Proteins 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 102000003946 Prolactin Human genes 0.000 description 1
- 108010057464 Prolactin Proteins 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 102000007327 Protamines Human genes 0.000 description 1
- 108010007568 Protamines Proteins 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 241000588770 Proteus mirabilis Species 0.000 description 1
- 241000588767 Proteus vulgaris Species 0.000 description 1
- 108010094028 Prothrombin Proteins 0.000 description 1
- 102100027378 Prothrombin Human genes 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 102000003743 Relaxin Human genes 0.000 description 1
- 108090000103 Relaxin Proteins 0.000 description 1
- 241000702263 Reovirus sp. Species 0.000 description 1
- 241000725643 Respiratory syncytial virus Species 0.000 description 1
- 241000235527 Rhizopus Species 0.000 description 1
- 240000005384 Rhizopus oryzae Species 0.000 description 1
- 235000013752 Rhizopus oryzae Nutrition 0.000 description 1
- 241000235546 Rhizopus stolonifer Species 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- 241000606723 Rickettsia akari Species 0.000 description 1
- 241000606720 Rickettsia australis Species 0.000 description 1
- 241000606695 Rickettsia rickettsii Species 0.000 description 1
- 241000606726 Rickettsia typhi Species 0.000 description 1
- 241000711897 Rinderpest morbillivirus Species 0.000 description 1
- 241000710799 Rubella virus Species 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241001138501 Salmonella enterica Species 0.000 description 1
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 1
- 241000223598 Scedosporium boydii Species 0.000 description 1
- 108010086019 Secretin Proteins 0.000 description 1
- 102100037505 Secretin Human genes 0.000 description 1
- 241000607766 Shigella boydii Species 0.000 description 1
- 241000607762 Shigella flexneri Species 0.000 description 1
- 241000607760 Shigella sonnei Species 0.000 description 1
- 241000710960 Sindbis virus Species 0.000 description 1
- 208000001203 Smallpox Diseases 0.000 description 1
- 241000589970 Spirochaetales Species 0.000 description 1
- 241001085826 Sporotrichum Species 0.000 description 1
- 241000295644 Staphylococcaceae Species 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 241000191963 Staphylococcus epidermidis Species 0.000 description 1
- 241000193998 Streptococcus pneumoniae Species 0.000 description 1
- 241000193996 Streptococcus pyogenes Species 0.000 description 1
- 241000194024 Streptococcus salivarius Species 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 102000011923 Thyrotropin Human genes 0.000 description 1
- 108010061174 Thyrotropin Proteins 0.000 description 1
- 102100028709 Thyroxine-binding globulin Human genes 0.000 description 1
- 108010011095 Transcortin Proteins 0.000 description 1
- 102000014034 Transcortin Human genes 0.000 description 1
- 108020004566 Transfer RNA Proteins 0.000 description 1
- 102000004338 Transferrin Human genes 0.000 description 1
- 108090000901 Transferrin Proteins 0.000 description 1
- 241000045663 Trematosphaeria grisea Species 0.000 description 1
- 241000589884 Treponema pallidum Species 0.000 description 1
- 241000589904 Treponema pallidum subsp. pertenue Species 0.000 description 1
- 241001045770 Trichophyton mentagrophytes Species 0.000 description 1
- 241000223229 Trichophyton rubrum Species 0.000 description 1
- 102000004903 Troponin Human genes 0.000 description 1
- 108090001027 Troponin Proteins 0.000 description 1
- 206010046865 Vaccinia virus infection Diseases 0.000 description 1
- 206010046980 Varicella Diseases 0.000 description 1
- 241000870995 Variola Species 0.000 description 1
- 241000700647 Variola virus Species 0.000 description 1
- GXBMIBRIOWHPDT-UHFFFAOYSA-N Vasopressin Natural products N1C(=O)C(CC=2C=C(O)C=CC=2)NC(=O)C(N)CSSCC(C(=O)N2C(CCC2)C(=O)NC(CCCN=C(N)N)C(=O)NCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(CCC(N)=O)NC(=O)C1CC1=CC=CC=C1 GXBMIBRIOWHPDT-UHFFFAOYSA-N 0.000 description 1
- 108010004977 Vasopressins Proteins 0.000 description 1
- 102000002852 Vasopressins Human genes 0.000 description 1
- 241000607626 Vibrio cholerae Species 0.000 description 1
- 102000050760 Vitamin D-binding protein Human genes 0.000 description 1
- 101710179590 Vitamin D-binding protein Proteins 0.000 description 1
- 241000710951 Western equine encephalitis virus Species 0.000 description 1
- 241000710772 Yellow fever virus Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000007801 affinity label Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229940005347 alcaligenes faecalis Drugs 0.000 description 1
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 1
- 102000013529 alpha-Fetoproteins Human genes 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- ORWYRWWVDCYOMK-HBZPZAIKSA-N angiotensin I Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C1=CC=C(O)C=C1 ORWYRWWVDCYOMK-HBZPZAIKSA-N 0.000 description 1
- 238000005571 anion exchange chromatography Methods 0.000 description 1
- 230000000603 anti-haemophilic effect Effects 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
- KBZOIRJILGZLEJ-LGYYRGKSSA-N argipressin Chemical compound C([C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSC[C@@H](C(N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N1)=O)N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(N)=O)C1=CC=CC=C1 KBZOIRJILGZLEJ-LGYYRGKSSA-N 0.000 description 1
- 229910052786 argon Inorganic materials 0.000 description 1
- 229940091771 aspergillus fumigatus Drugs 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 229940065181 bacillus anthracis Drugs 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000029918 bioluminescence Effects 0.000 description 1
- 238000005415 bioluminescence Methods 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- QXZGBUJJYSLZLT-FDISYFBBSA-N bradykinin Chemical compound NC(=N)NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(=O)NCC(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CO)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)CCC1 QXZGBUJJYSLZLT-FDISYFBBSA-N 0.000 description 1
- 229940056450 brucella abortus Drugs 0.000 description 1
- 229940038698 brucella melitensis Drugs 0.000 description 1
- 229960004015 calcitonin Drugs 0.000 description 1
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 1
- 229940095731 candida albicans Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 235000011089 carbon dioxide Nutrition 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- AOXOCDRNSPFDPE-UKEONUMOSA-N chembl413654 Chemical compound C([C@H](C(=O)NCC(=O)N[C@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@H](CCSC)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](C)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H]1N(CCC1)C(=O)CNC(=O)[C@@H](N)CCC(O)=O)C1=CC=C(O)C=C1 AOXOCDRNSPFDPE-UKEONUMOSA-N 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 229940048961 cholinesterase Drugs 0.000 description 1
- 229940015047 chorionic gonadotropin Drugs 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000005757 colony formation Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 108091007930 cytoplasmic receptors Proteins 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 206010013023 diphtheria Diseases 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- ZWIBGKZDAWNIFC-UHFFFAOYSA-N disuccinimidyl suberate Chemical compound O=C1CCC(=O)N1OC(=O)CCCCCCC(=O)ON1C(=O)CCC1=O ZWIBGKZDAWNIFC-UHFFFAOYSA-N 0.000 description 1
- 239000002532 enzyme inhibitor Substances 0.000 description 1
- 229940125532 enzyme inhibitor Drugs 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 102000034240 fibrous proteins Human genes 0.000 description 1
- 108091005899 fibrous proteins Proteins 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 238000002875 fluorescence polarization Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 229960002949 fluorouracil Drugs 0.000 description 1
- 229940028334 follicle stimulating hormone Drugs 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 230000007274 generation of a signal involved in cell-cell signaling Effects 0.000 description 1
- MASNOZXLGMXCHN-ZLPAWPGGSA-N glucagon Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 MASNOZXLGMXCHN-ZLPAWPGGSA-N 0.000 description 1
- 229960004666 glucagon Drugs 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000002622 gonadotropin Substances 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229960003786 inosine Drugs 0.000 description 1
- 108010093564 inter-alpha-inhibitor Proteins 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- SFVVQRJOGUKCEG-UHFFFAOYSA-N isoechinatine Natural products C1CC(O)C2C(COC(=O)C(O)(C(C)O)C(C)C)=CCN21 SFVVQRJOGUKCEG-UHFFFAOYSA-N 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 108010045069 keyhole-limpet hemocyanin Proteins 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 229940040129 luteinizing hormone Drugs 0.000 description 1
- 210000004880 lymph fluid Anatomy 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000006249 magnetic particle Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 208000020298 milker nodule Diseases 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 208000008588 molluscum contagiosum Diseases 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 125000004573 morpholin-4-yl group Chemical group N1(CCOCC1)* 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 238000002515 oligonucleotide synthesis Methods 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- XNOPRXBHLZRZKH-DSZYJQQASA-N oxytocin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSC[C@H](N)C(=O)N1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)NCC(N)=O)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 XNOPRXBHLZRZKH-DSZYJQQASA-N 0.000 description 1
- 229960001723 oxytocin Drugs 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 239000000199 parathyroid hormone Substances 0.000 description 1
- 229960001319 parathyroid hormone Drugs 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920001308 poly(aminoacid) Polymers 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000004800 polyvinyl chloride Substances 0.000 description 1
- 238000012987 post-synthetic modification Methods 0.000 description 1
- 230000001124 posttranscriptional effect Effects 0.000 description 1
- 230000001323 posttranslational effect Effects 0.000 description 1
- 150000003141 primary amines Chemical group 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 229940097325 prolactin Drugs 0.000 description 1
- 229940070353 protamines Drugs 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 229940007042 proteus vulgaris Drugs 0.000 description 1
- 229940039716 prothrombin Drugs 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 150000003230 pyrimidines Chemical class 0.000 description 1
- 238000000163 radioactive labelling Methods 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 108091092562 ribozyme Proteins 0.000 description 1
- 229940075118 rickettsia rickettsii Drugs 0.000 description 1
- 238000010850 salt effect Methods 0.000 description 1
- 229960002101 secretin Drugs 0.000 description 1
- OWMZNFCDEHGFEP-NFBCVYDUSA-N secretin human Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(N)=O)[C@@H](C)O)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)C1=CC=CC=C1 OWMZNFCDEHGFEP-NFBCVYDUSA-N 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 229940115939 shigella sonnei Drugs 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 210000001179 synovial fluid Anatomy 0.000 description 1
- 101150075675 tatC gene Proteins 0.000 description 1
- 210000001138 tear Anatomy 0.000 description 1
- 230000002277 temperature effect Effects 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 229960000874 thyrotropin Drugs 0.000 description 1
- 230000001748 thyrotropin Effects 0.000 description 1
- 239000012581 transferrin Substances 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000002753 trypsin inhibitor Substances 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001529453 unidentified herpesvirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 208000007089 vaccinia Diseases 0.000 description 1
- 229960003726 vasopressin Drugs 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 229940118696 vibrio cholerae Drugs 0.000 description 1
- 108700026220 vif Genes Proteins 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- 229940051021 yellow-fever virus Drugs 0.000 description 1
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
- C12Q1/6813—Hybridisation assays
- C12Q1/6832—Enhancement of hybridisation reaction
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
- C07K16/3069—Reproductive system, e.g. ovaria, uterus, testes, prostate
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1075—Isolating an individual clone by screening libraries by coupling phenotype to genotype, not provided for in other groups of this subclass
-
- 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
- C12Q1/6804—Nucleic acid analysis using immunogens
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
-
- 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
- C12Q2304/00—Chemical means of detecting microorganisms
- C12Q2304/10—DNA staining
-
- 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
- C12Q2527/00—Reactions demanding special reaction conditions
- C12Q2527/101—Temperature
-
- 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
- C12Q2537/00—Reactions characterised by the reaction format or use of a specific feature
- C12Q2537/10—Reactions characterised by the reaction format or use of a specific feature the purpose or use of
- C12Q2537/101—Homogeneous assay format, e.g. one pot reaction
-
- 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
- C12Q2563/00—Nucleic acid detection characterized by the use of physical, structural and functional properties
- C12Q2563/131—Nucleic acid detection characterized by the use of physical, structural and functional properties the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin
-
- 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
- C12Q2563/00—Nucleic acid detection characterized by the use of physical, structural and functional properties
- C12Q2563/179—Nucleic acid detection characterized by the use of physical, structural and functional properties the label being a nucleic acid
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2458/00—Labels used in chemical analysis of biological material
- G01N2458/10—Oligonucleotides as tagging agents for labelling antibodies
Definitions
- This invention relates to methods, compositions and kits for determining the presence or concentration of an analyte in a sample.
- Prostate specific antigen is a serum protein useful in the detection of prostate disease that is normally present in males at concentrations of about 0-4 ng/ml. PSA levels above 4 ng/ml are suspicious for prostate disease, particularly prostate cancer. This concentration range is readily detected by conventional immunoassay technology. Following prostate removal, however, the concentration of PSA drops to levels that are undetectable by conventional technology. Increasing PSA levels in cancer patients that have undergone prostate removal is indicative of relapse. An assay with femtogram/ml sensitivity is required to monitor these patients.
- Homogeneous immunoassays have been described for small molecules, such as drugs. These assays include SYVA's FRAT ® assay, EMIT ® assay, enzyme channeling immunoassay, and fluorescence energy transfer immunoassay (FETI) (Dade Behring, Deerfield, Illinois); enzyme inhibitor immunoassays (Hoffman LaRoche and Abbott Laboratories): fluorescence polarization immunoassay (Dandlicker), among others. All of these methods have limited sensitivity, and only a few, including FETI and enzyme channeling, are suitable for large multiepitopic analytes. Thus, there exists a need for a sensitive, homogeneous method for the detection of large and/or complex analytes present in biological and clinical samples.
- the present invention provides a binding pair having a first binding member comprising a first specificity molecule coupled to a first nucleic acid, and a second binding member comprising a second specificity molecule coupled to a second nucleic acid, where the first and second nucleic acids form a duplex of defined and limited stability.
- the first and second specificity molecules may be receptors, ligands, or antibodies.
- the specificity molecules may be identical or different from each other, hi one embodiment, the specificity molecules of the present invention interact with two receptors on a single cell.
- both the first and second specificity molecules are monoclonal antibodies, which may interact with different epitopes on the same antigen and thereby comprise a sandwich pair.
- the first and second nucleic acids of the binding pairs are typically single-strand nucleic acids which may be DNA, RNA, or PNA, but may be partially double-stranded nucleic acids or analogues thereof. In certain embodiments of the invention, at least one of the nucleic acids is a chimeric DNA/RNA molecule.
- the nucleic acids of the invention may be coupled via their 5' ends or their 3' ends. In one aspect of the invention, one nucleic acid of a binding pair is coupled via its 5' end and the other via its 3' end.
- the duplex between the nucleic acids may be formed between terminal ends of the nucleic acids or may comprise an internal nucleic acid sequence.
- the nucleic acid is suitable for amplification by PCR, LCR, SDA, or TMA.
- the present invention also provides methods for detecting an analyte using binding members.
- a binding pair as described above, is contacted with an analyte to form a complex.
- the binding pair nucleic acid is then dissociated and then re-associated.
- the reformed duplex may be detected, typically by amplifying a nucleic acid comprising the duplex by PCR. Background can be reduced significantly when the PCR primers bind only to sites generated by extending the 3' ends of the reformed duplex but not to the nucleic acids of the binding members themselves.
- the amplification products can be detected by any one of a variety of methods including staining with ethidium bromide, silver staining, autoradiography, dot blotting, slot blotting, southern blotting, and incorporation of a fluorescent molecule, a fluorescence quencher molecule, a chemiluminescent compound, a chemiluminescence quencher molecule, a bioluminescent compound or a fluorescent nucleotide.
- one of the binding pair nucleic acids is a chimeric DNA/RNA molecule and the other is a DNA molecule, where the duplex formed between the two nucleic acids has a short DNA/DNA hybrid region and longer DNA/RNA hybrid region.
- the intact duplex of this embodiment is stable, but can be destabilized by digestion with RNAse, which further reduces the background due to binding members that are not bound to analyte.
- FIG. 1 illustrates the content, conformation and general binding scheme for binding pairs.
- FIG. 2 provides a general scheme for duplex formation, 3' extension and PCR amplification of a nucleic acid pair comprising SEQ ID NO.:1 and SEQ ID NO.:2.
- FIG. 3 provides a general scheme for duplex formation, 3' extension and PCR amplification of a nucleic acid pair comprising SEQ ID NO.:1 and SEQ ID NO.:3.
- FIG. 4 shows the real-time PCR amplification (hot start) of both the 9 and 15 base pair overlapping oligonucleotide strands.
- FIG. 5 shows a schematic representation of a homogeneous NADIATM.
- FIG. 6 shows the orientation and duplex formation of a nucleic acid pair where one of the oligonucleotides [SEQ ID NO.:1] is attached via a spacer attached to its 3' end and the other oligonucleotide SEQ ID NO/.3] is attached directly via its 5' end.
- FIG 7. illustrates the process for use of chimeric RNA/DNA oligonucleotides in analyte detection.
- R indicates the position of ribonucleotide bases
- D indicates the position of deoxyribonucleotide bases
- S indicates the position of spacer molecules.
- FIG 8. shows a graph of the detection of PSA by homogeneous NADIATM
- Standard techniques may be used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, delivery, and treatment of patients. Standard techniques may be used for recombinant DNA methodology, oligonucleotide synthesis, tissue culture and the like. Reactions and purification techniques may be performed e.g., using kits according to manufacturer's specifications, as commonly accomplished in the art or as described herein.
- Binding member refers to a conjugate formed between a specificity molecule and a nucleic acid.
- compositions containing two binding members linked by a duplex of defined and limited stability formed between the binding member nucleic acids are referred to as "binding pairs.” Binding pairs combine with an analyte to form a binding pair-analyte complex which is herein referred to simply as a "complex.”
- analyte refers to any substance that it is desirable to detect in an assay, and which may be present in a sample.
- the analyte can be, without limitation, any substance.
- an analyte comprises a substance for which there exists a naturally occurring antibody or for which an antibody can be prepared.
- the analyte may be, for example, a protein, a polypeptide, a hapten, a carbohydrate, a lipid, a drug, a cell or any other of a wide variety of biological or non-biological molecules, complexes or combinations thereof.
- the analyte is a nucleic acid.
- the analyte is an antibody.
- the analyte is a cell (animal, plant, fungal, bacterial, etc.) or a subcomponent or organelle (e.g., mitochondria) thereof.
- Polyvalent ligand analytes that can be detected using compositions, methods and kits of the present invention will normally be poly(amino acids), i.e., polypeptides, proteins, polysaccharides, nucleic acids, and combinations thereof. Such combinations include components of cells, tissues, bacteria, viruses, cell walls, cell membranes, cellular organelles, chromosomes, genes, mitochondria, nuclei and the like. According to one aspect of the invention, certain analytes do not contain nucleic acid. A wide variety of protein analytes may be advantageously detected using the methods of the present invention.
- Such protein analytes can be classified according to family, with each family having similar structural features, biological functions, relationship to specific microorganisms (particularly disease causing microorganisms), and the like.
- Protein families of particular interest for the present invention include, for example, immunoglobulins, cytokines, enzymes, hormones, cancer antigens, nutritional markers, tissue specific antigens, and biowarfare agents. These protein analytes may be present in blood, serum, plasma, spinal fluid, synovial fluid, saliva, urine, cells or tissues.
- compositions, methods and kits of the present invention protamines histones albumins globulins scleroproteins phosphoproteins mucoproteins
- compositions, methods and kits of the present invention are clinically important proteins found in human plasma that may be detected using the compositions, methods and kits of the present invention: ⁇ i -Lipoprotein ⁇ l -Antitrypsin
- Hp 2-2 Ceruloplasmin Cholinesterase (X 2 -Lipoprotein(s) Myoglobin C-Reactive Protein ⁇ 2 -Macroglobulin ⁇ 2 -HS-glycoprotein Zn- ⁇ 2 -glycoprotein ⁇ 2 -Neuramino-glycoprotein Erythropoietin ⁇ -lipoprotein Transferrin Hemopexin Fibrinogen Plasminogen ⁇ 2 -glycoprotein I ⁇ 2 -glycoprotein II Immunoglobulin G (IgG) or ⁇ G-globulin
- compositions, methods and kits of the present invention include the examples listed in the Table below.
- compositions, methods and kits of the present invention include:
- Parathyroid hormone parathromone
- Melanotropin (melanocyte-stimulating hormone; intermedin)
- Luteinizing hormone interstitial cell-stimulating hormone
- Gonadotropin chorionic gonadotropin
- Bacteria and viruses are also analytes that may be detected using the compositions, methods and kits of the present invention. Included among these biological analytes are, among others: Corynebacteria
- Streptococcus pyrogenes Streptococcus salivarus
- Salmonella typhosa Salmonella typhosa
- Chlamydia (unclassifiable parasitesbacterial/viral) Chlamydia agents (naming uncertain)
- HIV Human Immunodeficiency Viruses I and II
- HTLV Human T-cell Lymphotrophic Virus I& II
- the endogenous cells of a human patient are analytes that may be advantageously detected using the compositions, methods and kits of the present invention.
- sample refers to an aliquot of material, frequently an aqueous solution or an aqueous suspension derived from biological material.
- Samples to be assayed for the presence of an analyte by the methods of the present invention include, for example, cells, tissues, homogenates, lysates, extracts, purified or partially purified proteins and other biological molecules and mixtures thereof.
- samples typically used in the methods of the invention include human and animal body fluids such as whole blood, serum, plasma, cerebrospinal fluid, sputum, bronchial washings, bronchial aspirates, urine, lymph fluids and various external secretions of the respiratory, intestinal and genitourinary tracts, tears, saliva, milk, white blood cells, myelomas and the like; biological fluids such as cell culture supernatants; tissue specimens which may or may not be fixed; and cell specimens which may or may not be fixed.
- human and animal body fluids such as whole blood, serum, plasma, cerebrospinal fluid, sputum, bronchial washings, bronchial aspirates, urine, lymph fluids and various external secretions of the respiratory, intestinal and genitourinary tracts, tears, saliva, milk, white blood cells, myelomas and the like
- biological fluids such as cell culture supernatants
- tissue specimens which may or may not be fixed and cell specimens which may or may not be fixed
- samples used in the methods of the present invention will vary based on the assay format and the nature of the tissues, cells, extracts or other materials, especially biological materials, to be assayed.
- Methods for preparing e.g. homogenates and extracts, such as protein extracts, from cells or other samples are well known in the art and can be readily adapted in order to obtain a sample that is compatible with the methods of the invention.
- contacting refers generally to providing access of one component, reagent, analyte or sample to another.
- contacting can involve mixing a solution comprising a binding pair with a sample comprising an analyte.
- the solution comprising one component, reagent, analyte or sample may also comprise another component or reagent, such as dimethyl sulfoxide (DMSO) or a detergent, which facilitates mixing, interaction, uptake, or other physical or chemical phenomenon advantageous to the contact between components, reagents, analytes and/or samples.
- DMSO dimethyl sulfoxide
- a detergent such as sodium sulfoxide
- contacting involves adding a solution comprising a binding pair to a sample comprising an analyte utilizing a delivery apparatus, such as a pipette-based device or syringe-based device.
- detecting refers to any method of verifying the presence of a given molecule.
- the techniques used to accomplish this may include, but are not limited to, PCR, nucleotide sequencing, PCR sequencing, molecular beacon technology, hybridization, hybridization followed by PCR, fluorescence, radiolabelling, phosphorescence and absorbance.
- reagents that may be used for detection include, but are not limited to, radiolabels, enzymatic labels (e.g. horseradish peroxidase, alkaline phosphatase), fluorescence, phosphorescence, bioluminescence, chemiluminescence, affinity labels (e.g. biotin, avidin, or streptavidin) and other reagents well known by those of skill in the art.
- the term "distinguishable” refers to an ability to distinguish experimentally between different markers, species or analytes.
- a binding pair or assay of the present invention may be used to detect multiple analytes.
- none of the nucleic acid markers will consist of sequences identical in both length and sequence.
- Two markers may comprise the same core sequence, but the markers will be distinguishable on the basis of size and/or different sequences.
- the detection products will be different in length.
- the markers will have sequences which bind to different sequence-specific probes.
- polynucleotide and “nucleic acid (molecule)" are used interchangeably to refer to polymeric forms of nucleotides of any length.
- the polynucleotides may contain deoxyribonucleotides, ribonucleotides and/or their analogs. Nucleotides may have any three-dimensional structure, and may perform any function, known or unknown.
- polynucleotide includes single-stranded, double- stranded and triple helical molecules.
- Olionucleotide refers generally to polynucleotides of between 5 and about 100 nucleotides of single- or double-stranded nucleic acid, typically DNA.
- Oligonucleotides are also known as oligomers or oligos and may be isolated from genes, or synthesized (e.g., chemically or enzymatically) by methods known in the art.
- a "primer” refers to an oligonucleotide, usually single- stranded, that provides a 3'-hydroxyl end for the initiation of enzyme-mediated nucleic acid synthesis.
- polynucleotides a gene, a gene fragment, exons, introns, mRNA, tRNA, rRNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes and primers.
- a nucleic acid molecule may also comprise modified nucleic acid molecules, such as methylated nucleic acid molecules and nucleic acid molecule analogs.
- Analogs of purines and pyrimidines are known in the art, and include, but are not limited to, aziridinycytosine, 4- acetylcytosine, 5-fluorouracil, 5-bromouracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethyl-aminomethyluracil, inosine, N6-isopentenyladenine, 1-methyladenine, 1-methylpseudouracil, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2- methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, pseudouracil, 5- pentylnyluracil and 2,6-diaminopurine.
- uracil as a substitute for thymine in a deoxyribonucleic acid is also considered an analogous form of pyrimidine.
- nucleic acid molecules of the present invention may comprise a modified sugar and a modified phosphate backbone.
- a nucleic acid of the invention may comprise modifications to sugar, base and phosphate backbone.
- nucleic acid marker refers to a nucleic acid molecule that will produce a detection product of a predicted size or other selected characteristic when used with appropriately designed oligonucleotide primers in a nucleic acid amplification reaction, such as a PCR reaction. Skilled artisans will be familiar with the design of suitable oligonucleotide primers for PCR and programs are available commercially and over the Internet to facilitate this aspect of the invention ⁇ see, for example, the http site: bibiserv.techfak.unibielefeld.de/genefisher).
- a nucleic acid marker may be linear or circular.
- the nucleic acid marker will comprise a predetermined, linear nucleic acid sequence with binding sites for selected primers located at or near each end.
- the primers will be internal rather than at an end, and a single primer may be used, e.g. for Rolling Circle Amplification.
- Amplified DNA may be detected using any available method, including, but not limited to techniques such as real-time PCR, SYBR ® Green staining, or ethidium bromide staining.
- the binding sites for the amplification primers flank an undefined DNA sequence of defined length, or a DNA sequence that comprises another identifiable characteristic, such as a detectable sequence, in addition to undefined sequences.
- the nucleic acid marker is distinguished by the size or mass of the amplified sequences; thus, the DNA sequence between the primers need not be defined as to the exact sequence, just as to the number of bases. Alternatively, the size and/or sequence of the entire nucleic acid marker need not be defined as long as a binding site for a molecular beacon ⁇ see, infra) is supplied. In further embodiments, the DNA sequence located between the primer binding sites comprises a "characteristic identification sequence" capable of being detected during the PCR reaction.
- Fluorescent signal generation may, for example, be sequence-specific (Molecular Beacons, TaqMan ® , fiuorogenic primers, such as the LUXTM primers (Invitrogen (Carlsbad, CA.)) or mass dependent ⁇ e.g., SYBR ® Green, Ethidium Bromide).
- sequence-specific primers such as the LUXTM primers (Invitrogen (Carlsbad, CA.)
- mass dependent ⁇ e.g., SYBR ® Green, Ethidium Bromide mass dependent ⁇ e.g., SYBR ® Green, Ethidium Bromide.
- the term "specificity molecule” as used herein refers to any molecule that is capable of specifically binding another molecule. In one embodiment, the specificity molecule is an antibody.
- specificity molecules can include, without limitation: biological receptor molecules, such as the insulin receptor; ligands for receptors, (e.g., insulin for the insulin receptor); antigens (e.g., to bind to antibodies) and biological, chemical or other molecules that have affinity for another molecule, such as biotin and avidin.
- biological receptor molecules such as the insulin receptor
- ligands for receptors e.g., insulin for the insulin receptor
- antigens e.g., to bind to antibodies
- biological, chemical or other molecules that have affinity for another molecule such as biotin and avidin.
- the specificity molecules of the present invention need not comprise an entire naturally occurring molecule but may consist of only a portion, fragment or subunit of a naturally occurring molecule, as for example the Fab fragment of an antibody.
- Specificity molecules may be generated by any method known in the art.
- antibodies may be found in an antiserum, prepared from a hybridoma tissue culture supernatant or ascites fluid, or may be derived from a recombinant expression system, as will be well known in the art. Fragments, portions or subunits of e.g., an antibody, receptor or other species, may be generated by chemical, enzymatic or other means, yielding for example, well-known (e.g., Fab, Fab') or novel molecules.
- specificity molecules can include recombinant, chimeric and hybrid molecules, such as humanized and primatized antibodies, and other non-naturally occurring antibody forms.
- binding and “specific binding” as used herein is meant that an antibody or other molecule, especially a specificity molecule of the invention, binds to a target such as an antigen, ligand or other analyte, with greater affinity than it binds to other molecules under the specified conditions of the present invention.
- Antibodies or antibody fragments as known in the art, are polypeptide molecules that contain regions that can bind other molecules, such as antigens.
- “specifically binding” may mean that an antibody or other specificity molecule binds to a target analyte molecule with at least about a 10 6 -fold greater affinity, preferably at least about a 10 7 -fold greater affinity, more preferably at least about a 10 8 -fold greater affinity, and most preferably at least about a 10 9 -fold greater affinity than it binds molecules unrelated to the target molecule.
- specific binding refers to affinities in the range of about 10 6 -fold to about 10 9 -fold greater than non-specific binding, hi some embodiments, specific binding may be characterized by affinities greater than 10 9 -fold over non-specific binding.
- Polyclonal Antibodies are heterogeneous populations of antibody molecules derived from the sera of animals immunized with an antigen, or an antigenic functional derivative thereof.
- host animals such as rabbits, mice and goats, may be immunized by injection with an antigen or hapten-carrier conjugate, optionally supplemented with adjuvants.
- Polyclonal antibodies may be unpurified, purified or partially purified from other species in an antiserum.
- “Monoclonal antibodies,” or “MAbs,” which are homogeneous populations of antibodies to a particular antigen, may be obtained by any technique that provides for the production of antibody molecules, such as by continuous culture of cell lines. These techniques include, but are not limited to the hybridoma technique of K ⁇ hler and Milstein, Nature, 256:495-7 (1975); and U.S. Patent No. 4,376,110), the human B-cell hybridoma technique (Kosbor, et ah, Immunology Today, 4:72 (1983); Cote, et ah, Proc. Natl. Acad. Sci.
- Such antibodies may be of any immunoglobulin class including IgG, IgM, IgE, IgA, IgD and any subclass thereof.
- the hybridoma producing the MAb of this invention may be cultivated in vitro or in vivo. Production of high titers of MAbs in vivo makes this a presently preferred method of production.
- chimeric antibodies In addition, techniques developed for the production of "chimeric antibodies" (Morrison, et ah, Proc. Natl. Acad. ScL, 81:6851-6855 (1984); Takeda, et ah, Nature, 314:452-54 (1985)) by splicing the genes from a mouse antibody molecule of appropriate antigen specificity together with genes from a human antibody molecule of appropriate biological activity can be used.
- a chimeric antibody can be a molecule in which different portions are derived from different animal species, such as those having a variable region derived from a murine MAb and a human immunoglobulin constant region.
- Single chain antibodies are typically formed by linking the heavy and light chain fragments of the Fv region via an amino acid bridge, resulting in a single chain polypeptide.
- Antibody fragments that recognize specific epitopes may be generated by known techniques.
- such fragments include but are not limited to: the F(ab') 2 fragments that can be produced by pepsin digestion of the antibody molecule and the Fab fragments that can be generated by reducing the disulfide bridges of the F(ab') 2 fragments.
- Fab expression libraries may be constructed (Huse, et ah, Science, 246:1275-81 (1989)) to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity.
- hapten refers to a small proteinaceous or non-protein antigenic determinant which is capable of being recognized by an antibody. Typically, haptens do not elicit antibody formation in an animal unless part of a larger species. For example, small peptide haptens are frequently coupled to a carrier protein, such as keyhole limpet hemocyanin, in order to generate an anti-hapten antibody response.
- Antigens are macromolecules capable of generating an antibody response in an animal and being recognized by the resulting antibody. Both antigens and haptens comprise at least one antigenic determinant or "epitope," which is the region of the antigen or hapten which binds to the antibody. Typically, the epitope on a hapten is the entire molecule.
- sandwich pair antibodies or "sandwich antibody pairs,” as used herein, refers to a pair of typically monospecific antibodies, e.g. monoclonal antibodies, that are suitable for use in a sandwich format immunoassay. Each antibody of the pair binds to a different epitope on the same molecule and both antibodies of the pair can bind to the antigen at the same time. Methods for identifying pairs of antibodies suitable for sandwich assays will be well known to those in the art. The skilled artisan will also recognize that various other molecules can be used as sandwich pairs. For example, a receptor analyte can be sandwiched between a ligand for that receptor and an antibody that binds to an epitope on the receptor that is not involved in ligand binding. Thus, an antibody and ligand can be used as a sandwich pair for a receptor analyte.
- Receptor typically refers to a molecular structure within or on the surface a cell characterized by selective binding of a specific substance (e.g. a "ligand") and resulting in a specific physiologic effect that accompanies the binding.
- a specific substance e.g. a "ligand”
- receptors include cell surface receptors for peptide hormones, neurotransmitters, antigens, complement fragments, immunoglobulins and cytoplasmic receptors for steroid hormones.
- the receptor will typically be isolated and purified and need not effect or be capable of effecting a physiological or other biological effect.
- the methods of the present invention exploit the selective binding of the receptor to the specific substance.
- ligand refers generally to a molecule that binds to a receptor.
- a ligand is a small, soluble molecule, such as a ho ⁇ none or neurotransmitter.
- solid support refers to any solid phase that can be used to immobilize e.g., an analyte, an antibody or a complex.
- Suitable solid supports will be well known in the art and include the walls of wells of a reaction tray, such as a microtiter plate, the walls of test tubes, polystyrene beads, paramagnetic or non-magnetic beads, nitrocellulose membranes, nylon membranes, microparticles such as latex particles, and sheep (or other animal) red blood cells.
- Typical materials for solid supports include, but are not limited to, polyvinyl chloride (PVC), polystyrene, cellulose, nylon, latex and derivatives thereof.
- the solid support may be coated, derivatized or otherwise modified to promote adhesion of the desired molecules (e.g., analytes) and/or to deter non-specific binding or other undesired interactions.
- desired molecules e.g., analytes
- deter non-specific binding or other undesired interactions e.g., analytes
- the choice of a specific "solid phase" is usually not critical and can be selected by one skilled in the art depending on the assay employed.
- latex particles, microparticles, paramagnetic or non-magnetic beads, membranes, plastic tubes, walls of microtiter wells, glass or silicon chips, and red blood cells are all suitable sold supports.
- the solid support can be selected to accommodate various detection methods.
- 96 or 384 well plates can be used for assays that will be automated, for example by robotic workstations, and/or those that will be detected using, for example, a plate reader.
- the solid support may be a thin membrane, such as a nitrocellulose or nylon membrane.
- sandwich immunoassays the walls of the wells of a reaction tray are typically employed.
- paramagnetic beads may be used as a solid support. Suitable methods for immobilizing molecules on solid phases include ionic, hydrophobic, covalent interactions and the like, and combinations thereof.
- a solid support as used herein may thus refer to any material that is insoluble, or can be made insoluble by a subsequent reaction.
- the solid support can be chosen for its intrinsic ability to attract and immobilize a capture reagent.
- the solid phase can retain an additional receptor that has the ability to attract and immobilize a capture reagent.
- the additional receptor may include a substance that is oppositely charged with respect to either the capture reagent itself or to a charged substance conjugated to the capture reagent.
- an additional receptor molecule can be any specific binding member that is immobilized upon (attached to) the solid phase and which has the ability to immobilize a capture reagent through a specific binding reaction.
- the additional receptor molecule enables indirect immobilization of the capture reagent to a solid phase before or during the performance of the assay.
- the solid phase thus can be a plastic, derivatized plastic, paramagnetic or non-magnetic metal, glass or silicon surface of a test tube, microtiter well, sheet, bead, microparticle, chip, or other configurations known to those of ordinary skill in the art.
- Protein generally refers to a short chain of amino acids linked by peptide bonds. Typically peptides comprise amino acid chains of about 2-100, more typically about 4-50, and most commonly about 6-20 amino acids.
- Polypeptide generally refers to individual straight or branched chain sequences of amino acids that are typically longer than peptides. Polypeptides usually comprise at least about 100 to 1000 amino acids in length, more typically at least about 150 to 600 amino acids, and frequently at least about 200 to about 500 amino acids.
- Proteins include single polypeptides as well as complexes of multiple polypeptide chains, which may be the same or different.
- Multiple chains in a protein may be characterized by secondary, tertiary and quaternary structure as well as the primary amino acid sequence structure; may be held together, for example, by disulfide bonds; and may include post-synthetic modifications such as, without limitation, glycosylation, phosphorylation, truncations or other processing.
- Antibodies such as IgG proteins are typically comprised of four polypeptide chains ⁇ i.e., two heavy and two light chains) that are held together by disulfide bonds.
- proteins may include additional components such associated metals ⁇ e.g., iron, copper and sulfur), or other moieties.
- peptides, polypeptides and proteins include, without limitation, biologically active and inactive forms; denatured and native forms; as well as variant, modified, truncated, hybrid, and chimeric forms thereof.
- the peptides, polypeptides and proteins of the present invention may be derived from any source or by any method, including, but not limited to extraction from naturally occurring tissues or other materials; recombinant production in host organisms such as bacteria, fungi, plant, insect or animal cells; and chemical synthesis using methods that will be well known to the skilled artisan.
- conjugate refers to two molecules that have been covalently attached or otherwise linked together.
- a nucleic acid conjugate is generated by covalently linking a nucleic acid to a protein, polypeptide or other specificity molecule.
- the protein, polypeptide or other specificity molecule is covalently attached to a nucleic acid via a linking group to form a conjugate.
- a "kit” for detecting the presence of an analyte in a sample by the methods of the invention may, by way of example, comprise at least one container means having disposed therein a binding pair specific for the selected analyte.
- the kit may further comprise other container means comprising one or more of the following: buffers, solutions or other reagents and materials necessary for performing analyte detection; reagents capable of amplifying the nucleic acid probe components of the binding pairs; and reagents capable of detecting the presence of nucleic acid components following amplification.
- the kit further comprises instructions for use.
- the kit if intended for diagnostic use, may also include notification of a FDA approved use and instructions therefor.
- a compartmentalized kit includes any kit in which reagents are contained in separate containers.
- Such containers include small glass containers, plastic containers or strips of plastic or paper.
- Such containers allow the efficient transfer of reagents from one compartment to another compartment such that the samples and reagents are not cross-contaminated and the agents or solutions of each container can be added in a quantitative fashion from one compartment to another.
- Such containers may include a container which will accept a test sample, a container which contains the probe or primers used in the assay, containers which contain buffers and reagents (such as phosphate buffered saline, Tris-buffers, and the like), and containers which contain the reagents used to detect the marker nucleic acid, amplified product, or the like.
- buffers and reagents such as phosphate buffered saline, Tris-buffers, and the like
- containers which contain the reagents used to detect the marker nucleic acid, amplified product, or the like.
- a kit for coupling DNA to an antibody or other specificity molecule by the methods of the invention may comprise at least one container means having disposed therein lyophilized, activated DNA.
- the kit may further comprise other containers comprising one or more of the following: reagents, buffers and agents capable of detecting the presence of nucleic acid after the reaction.
- the kit further comprises instructions for use.
- the activated nucleic acids described in the present invention can readily be incorporated into one of the established kit formats that are well known in the art.
- the present invention generally provides compositions, methods and kits for the use of novel nucleic acid labeled-binding pairs of defined and limited stability under defined conditions for the detection of analytes, particularly homogenous sensitive analyte detection. Binding Pairs
- the invention provides binding pairs that are useful for detecting analytes.
- Binding pairs of the present invention are formed between two binding members, each having a specificity molecule coupled to a nucleic acid, as shown in FIG. 1.
- the nucleic acid component of the binding members are complementary over at least a part of their length, and can form a duplex through which the two members of the binding pair are joined (see FIG. 1).
- the specificity molecules of each binding pair direct the interaction of the pair to a desired analyte.
- a specificity molecule of the invention can be any molecule that is capable of interacting with another molecule, such as an analyte.
- specificity molecules interact with an analyte with high specificity and affinity.
- the specificity molecule is a receptor. In another aspect, the specificity molecule is a ligand. In yet another aspect of the invention, the specificity molecule is an antibody.
- the individual specificity molecules of a binding pair can both be the same type of molecule or can be different types of molecules.
- both specificity molecules in a pair are receptors.
- both specificity molecules in pair are ligands.
- both are antibodies.
- both are antigens.
- one specificity molecule is a ligand and the other specificity molecule is a receptor.
- one or both of the specificity molecules in a binding pair is a nucleic acid.
- the target analyte is a complementary nucleic acid.
- the target analyte is a DNA, an RNA or a protein, or a component of a complex thereof.
- the target analyte is a complex comprising a nucleic acid and a protein and one of the specificity molecules of the binding pair is a nucleic acid that is complementary to the nucleic acid component of the target analyte and the other specificity molecule of the binding pair is a protein (i.e., an antibody, antigen, ligand, receptor, etc.) that binds specifically to the protein component of the target analyte.
- at least one specificity molecule of a binding pair is an antibody.
- Antibodies are particularly useful as specificity molecules. All types of antibodies described supra are useful for imparting specificity to a binding pair, including but not limited to monoclonal antibodies, polyclonal antibodies, chimeric antibodies, hybrid antibodies, recombinant antibodies, humanized antibodies, primatized antibodies, truncated antibodies, single chain antibodies and the like.
- the two antibodies When two antibodies are used to form a binding pair, the two antibodies may have the same or different specificities. In one embodiment of the invention, the two antibodies recognize the same epitope and that epitope is present multiple times on a single analyte. For example, both antibodies in a binding pair may recognize a receptor that is present in multiple copies on the surface of a cell analyte. In another embodiment, the antibodies recognize different epitopes on a single analyte molecule. In certain aspects of this embodiment, both antibodies can bind simultaneously to a single analyte molecule. The so called "sandwich pair" antibodies of this aspect of the invention will be well known to those skilled in the art. Nucleic Acid Components of Binding Pairs
- each specificity molecule in a binding pair is coupled to or "labeled" with a nucleic acid molecule ⁇ i.e., a nucleic acid marker).
- the nucleic acids are single-stranded.
- the nucleic acids are partially single-stranded.
- the nucleic acid is all DNA, all RNA or a mixture of DNA and RNA.
- nucleotide analogs or derivatives may be used.
- PNA is used as part or all of the nucleic acid.
- the specificity molecule may be coupled to either the 3 ' or 5' end of the nucleic acid.
- the terminal free ends of the two nucleic acids in a binding pair are complementary such that they can hybridize to form a duplex nucleic acid region of limited and defined stability.
- the terminal free ends that form a duplex are the 3' ends of the nucleic acids.
- the terminal free end of one of the nucleic acids of a binding pair is complementary to a sequence of the other nucleic acid of that binding pair at a point other than the terminal end of one of the nucleic acid.
- the 3' end of the first nucleic acid forms a duplex with an internal sequence of the second nucleic acid.
- the nucleic acid is coupled to a specificity molecule via one or more linkers or spacers.
- nucleotide sequence of the nucleic acids of the present invention is of less importance than the functional roles they are required to perform. Accordingly, the sequence of the nucleic acids, as well as the length of the nucleic acid component of the binding pair, may vary considerably, provided the nucleic acid component of the binding pair can still perform the functional roles they are required to perform. Importantly, the sequence and length of the nucleic acids of the binding pair are not limited to those exact sequences and lengths of the exemplary binding pairs disclosed herein. The nucleic acids of the binding pair thus can be of different lengths and or sequence. An important function of the nucleic acid component of the binding pairs of the present invention is to provide a highly sensitive label for analyte detection.
- the nucleic acid is amplified and detected as a measure of analyte concentration.
- the nucleic acid can be amplified using well established methods. Exemplary amplification methods include those described in various U.S. patents, the contents of which are incorporated by reference herein in their entirety: PCR ⁇ see e.g. U.S. Patent No. 4,683,202), TMA ⁇ see e.g. U.S. Patent No.5,399,491); SDA (see e.g. U.S. Patent No. 5,270,184), and LCR (see e.g. U.S. Patent No. 5,427,930).
- nucleic acid sequences can be used for amplification.
- the length of the nucleic acid can be varied considerably.
- the nucleic acids of the invention are designed to avoid intramolecular hairpin formation in order to increase the efficiency with which they can be amplified.
- the nucleic acid is detected using a nonspecific intercalating fluorescent dye, such as SYBR ® Green. According to this embodiment, the strength of the detection signal will be a function of the length of the nucleic acid detected.
- nucleic acid duplex is dependent in part on the length of the region of complementarity between the nucleic acid strands in the duplex. A longer complementarity region or "overlap" between nucleic acids increases the stability of the duplex that is formed. Conversely, a shorter overlap leads to a less stable duplex. Nucleic acids of the present invention are designed to have defined stability that can be manipulated by altering length, temperature, solvent and other conditions.
- Factors that influence the stability of the hybrid include, but are not limited to, the concentration of the nucleic acid-labeled binding pairs, salt concentration, temperature, organic solvents such as ethanol, DMSO, tetramethylammonium ions (TMA + ⁇ , base pair mismatches and the like.
- the nucleic acids are designed to form intra-binding pair duplexes.
- the nucleic acid duplexes of a binding pair can be dissociated under defined conditions and thus have limited stability under certain conditions.
- reaction conditions are designed to maximize duplex formation between the members of binding pairs bound to their respective target analytes while minimizing duplex formation between members of binding pairs that are not bound to target analytes.
- the duplexes remain stable during binding to an analyte, but can be selectively dissociated or melted by an assay operator.
- thermodynamic conditions favors the unbound pairs to be unhybridized.
- less than about 1% of the binding pairs are not bound to analyte.
- less than about 0.1%, 0.01%, or 0.001% of the binding pairs are not bound to analyte.
- a competitor nucleic acid e.g., complementary to a region of duplex formation between binding pair nucleic acids
- a similar effect may be obtained by designing one of the nucleic acids in a binding pair to form a small hairpin when it is not hybridized to the nucleic acid of its binding partner.
- the stability of the complex between the binding pair and the target analyte can be enhanced by using a polymerase enzyme to extend 3' ends of the nucleic acid duplex as described in U.S. Patent No. 5,635,602, the contents of which are incorporated by reference herein in its entirety.
- Nucleic acid markers of the present invention can be detected by incorporating any one of a number of labels into one or both of the binding pair nucleic acids.
- markers suitable for use in the present invention include intercalating fluorescent molecules such as SYBR ® Green and ethidium bromide; FRET fluorescent resonance energy transfer pairs, such as fluorescein and rhodamine; radioactive compounds, bioluminesceiit compounds, chemiluminescent compounds such as acridinium ester; chemiluminescent quencher pairs; and reagents that hybridize to nucleic acid in a sequence-specific manner, such as Molecular Beacons (Kramer), TaqMan ® , and fluorogenic primers (e.g.
- NTPs and dNTPs fluorescent nucleic acid triphosphates
- the present invention also provides methods for detecting analytes using binding pairs.
- an analyte-specific binding pair as described above, is contacted with its analyte, to form a complex.
- the members of the binding pair are joined by the intermolecular duplex of defined and limited stability formed between the two nucleic acids. Furthermore, the binding members are locked in place in close proximity, and therefore high local concentration, by virtue of their association with the analyte.
- the nucleic acid duplexes may then be dissociated and allowed to reassociate.
- dissociation may be accomplished by heating the complexes above the melting temperature of the nucleic acid duplex.
- the salt concentration or ionic strength may be decreased.
- a chemical or biological agent may be added to the complex to dissociate the duplexes.
- the reassociation of the nucleic acids of binding pair members not complexed with analyte may be prevented or eliminated by dilution.
- the dilution or other condition of the reaction does not allow substantial reassociation of excess free binding members.
- conditions which do not allow substantial reassociation involve less than 5%, typically less than 1%, frequently less than 0.1% and most often less than 0.01% reassociation of excess free binding members.
- binding pairs are contacted with an analyte such that a complex between the binding pair and the analyte is formed.
- the complexes are then diluted such that hybridization of proximity-bound nucleic acid markers is favored but hybridization in bulk solution is disfavored. Under these conditions, the extension of 3' ends of the marker nucleic acid occurs predominantly or exclusively in the complexed and hybridized marker nucleic acids, while the uncomplexed nucleic acids are not hybridized and therefore extension from 3' ends does not occur.
- the extended duplexes are then detected as a means for analyte detection.
- a nucleic acid comprising the duplex is amplified to facilitate detection. Amplification may be accomplished by any method known in art. Suitable methods include, but are not limited to PCR, LCR, SDA, and TMA.
- nucleic acid amplification methods rely on primed enzymatic synthesis of DNA.
- a primer with a free 3' OH is hybridized to a single- strand DNA or RNA template.
- the 3' OH can then form the initiation point for extension using a polymerase, examples of which are well known in the art and include, without limitation, Taq DNA polymerase, DNA Polymerase I, Klenow Fragment of DNA Polymerase, T4 DNA polymerase, T7 DNA Polymerase, Reverse Transcriptase, phi29 DNA Polymerase, Bst DNA Polymerase, AccuPrimeTM (Invitrogen), Pfic DNA Polymerase (Invitrogen), FideliTaqTM DNA Polymerase(Amersham Biosciences), SequenaseTM, Thermo SequenaseTM DNA, Hot TubTM DNA Polymerase (Amersham Biosciences), Vent ® Polymerase (New England Biolabs), and 9°N m DNA Polymerase.
- the template for PCR is a nucleic acid molecule comprising the duplex region of the two strands of nucleic acid complexed to the analyte via their respective specificity molecules.
- a template for PCR may be longer than the desired PCR product.
- the two individual PCR primers may hybridize internally and initiate DNA synthesis, each at a particular site on one strand of the nucleic acid template.
- the resulting PCR product typically has a 5' terminus defined by one primer and a 3' terminus defined by the other primer.
- the amplification products can then be detected, for example by staining with ethidium bromide, silver staining, autoradiography, dot blotting, slot blotting, or southern blotting.
- detection can be accomplished by incorporating a detection molecule into one or both of the nucleic acids, the amplification product or the duplex region of the complex.
- Detection molecules suitable for use in the methods of the present invention include, but are not limited to fluorescent molecules, fluorescence quencher molecules, chemiluminescent compound, chemiluminescence quencher molecules, a fluorescent nucleotides, enzymatic labels, and radiolabels.
- duplex detection may be facilitated by incorporating an acridinium ester into the duplex region of the complex and selectively hydrolyzing the label in first and second binding pair members that have not formed a nucleic acid duplex.
- the present invention provides methods for detecting analytes through the amplification of binding pair nucleic acid duplexes in which one or both primer binding sites are absent from nucleic acid marker sequences.
- one or more primer binding sites are generated after the binding pair has bound an analyte.
- one or both of the nucleic acid marker sequences may incorporate a sequence identical to the desired primer sequence, outside of the duplex region.
- the primer binding sites for amplification are not present in the binding pair nucleic acids but are only formed after the 3' ends of nucleic acid duplexes are extended.
- no region complementary to the primer i.e., no primer site
- the 3' end of one or both of the markers may be extended using a suitable polymerase.
- the extension encompasses the sequence identical to the desired primer sequence, thereby generating a complementary primer binding site which may be used in subsequent amplification steps for binding to the primer (FIGS. 2 and 3).
- the amplification primer will neither hybridize nor initiate nucleic acid synthesis/amplification unless a duplex region is formed between the first and second nucleic acid markers and subsequently extended.
- the background non-specific detection is reduced because only hybridized molecules are detected.
- Oligonucleotides 60 bases long having a 5' amino C 12 spacer arm (Spacer C 12 CE Phosphoramidite;12-(4,4'-Dimethoxytrityloxy)dodecyl-l-[(2-cyanoethyl)-(N,N- diisopropyl)]-phosphoramidite) with the following nucleotide sequences have been found suitable for use in this aspect of the of the present invention:
- Oligonucleotide (1) is complimentary to both oligonucleotide (2) [SEQ ID NO.:2] and oligonucleotide (3) [SEQ ID NO.:3] for the last 9 and 15 bases, respectively, at the 3' ends.
- the length of the duplex formed between the oligos has a specific Gibbs Free Energy ( ⁇ G), which is controlled by nearest neighbor base sequence. See Breslauer, et al, Proc. Nat 'I. Acad. Sci. U.S.A. 83: 3746-3750 (1986).
- the 9 and 15 base-pair overlaps have melting temperatures (T n ,' s) of 26° C and 42° C, respectively, in 50 niM NaCl according to the formula:
- T m 64.9 + 41 * (yG + zC - 16.4) / (wA + xT + yG + zC) where w, x, y, z are the number of the bases A, T, G, C in the sequence, respectively. See e.g., http://www.basic.nwu.edu/biotools/oligocalc.html.
- the overlapping strands hybridize (> 50% duplex) when present in solution at concentrations greater than 50 nM at 25° C, 50 niM NaCl.
- the strands exist predominately as monomers ( ⁇ 50% hybrid) at concentrations less than 100 pM at equilibrium.
- complementary DNA strands such as these may transition between single-and double-stranded forms as a function of temperature, DNA concentration and salt effects.
- the 3' end of each strand can serve as the initiation point for synthesis of nucleic acid complementary to the other strand.
- Each strand may be extended in the presence of an appropriate DNA polymerase and nucleotide triphosphates, resulting in a 111 (FIG. 2) or 105 (FIG. 3) base pair DNA duplex.
- the newly formed duplex contains sequences that were not present initially in the partially overlapped structure. These new sequences can be replicated exponentially by PCR in the presence of the following downstream primers:
- FIG. 4 shows the real-time PCR amplification (hot start) of both the 9 and 15 base pair overlapping oligonucleotide strands. It can be seen from this Figure that amplification does not occur until a sufficient concentration of strands are present to ensure the probability that one strand will be close enough to a complementary strand for hybridization and first chain extension. Increasing the concentration of overlapping strands results in an exponential increase in template generation, as measured by real-time PCR threshold cycle. Conversely, diluting this concentration, by washing a support for instance, exponentially decreases signal. Amplification of a normal 60-mer template is shown for comparison. Proximity Effects in Binding Pair-Analyte Complexes
- NADIATM Nucleic Acid Detection LnmunoAssay
- Antibody binding members labeled with overlapping oligonucleotides also exhibit the PCR amplification effect shown in FIG. 4. Binding such DNA-Ab conjugates to the same antigen, or to an antigen coated surface, fixes the overlapping oligonucleotide strands in proximity to one another. This effectively increases the relative concentrations of both strands and the probability of template formation. The template, once formed, exhibits a linear relationship between concentration and real-time PCR threshold signal.
- This proximity effect has implications for greatly reducing background signal arising from the nonspecific binding of template DNA-Ab conjugate when a capturing support is used (i.e. in a heterogeneous assay).
- the individual overlapping oligonucleotide sequences are not template molecules for amplification and, unlike DNA- Ab conjugates previously described, cannot be amplified by themselves with the selected primer set. Although the overlapping conjugates still bind nonspecifically, the binding is random. Random nonspecific binding of a relatively small number of copies of conjugate greatly diminishes the opportunity for proximal interaction and first chain extension.
- the proximity effect may also be the basis for homogeneous sandwich assay formats of the present invention, in which a capturing solid support is unnecessary.
- conditions must be used which favor the hybridization and first chain extension of proximally bound overlapping conjugates and disfavor hybridization in the bulk solution. Any nonspecific hybridization of the conjugate pair in solution will elicit signal, as shown in FIG. 4.
- One embodiment of the present invention therefore, entails employing the lowest conjugate concentration possible to maintain an acceptable immune complex formation rate. Adjustment of overlap length, salt conditions and temperature effects are also available to minimize random hybridization of the overlap sequences in the bulk solution. Immunodetection of Cellular Analytes
- Affinity purified polyclonal antibodies to E. coli 0157 have been conjugated to the overlapping oligonucleotide strands in order to demonstrate a heterogeneous NADIA format for the detection of intact microorganisms.
- Antibody attached to magnetic particles has been used to isolate and enrich E. coli 0157 cells from water samples and meat homogenate. The cell surface is estimated to exhibit several thousand copies of the specific antigen that was detected. Calculations show that the distance between randomly distributed sites fall within the spanning distance of overlapping oligonucleotides conjugated to anti-E. coli 0157. In this case, proximity binding is due to individual antigen spacing, as opposed to separate epitopes on a single protein antigen.
- polyclonal anti-E. coli 0157 (KPL, Gaithersburg, MD) was labeled in three separate reactions with sequences (1) (2) and (3) and maintained as individual conjugates.
- E. coli 0157:H7 was obtained from ATCC (700728) and grown at 37° C. Cells from liquid culture were diluted in fresh media and incubated with 10 nM overlapping oligonucleotide-Ab conjugates for 1 hour at room temperature. The resulting cell- antibody complex was washed free of excess, unbound conjugate by centrifugation and resuspension in cold tris-buffered saline, effectively diluting the overlapping antibody reagent to below 1.0 pM.
- the washed cells were simultaneously streaked onto plates and assayed by incubation in PCR reagent mixture for 10 min at 33° C, followed by 40 cycles of real-time PCR in the presence of downstream primers.
- the present invention provides a method for detecting intact cells or viruses by forming a complex on the cell surface with a binding pair reagent.
- the binding pair reagent used for detecting intact cells such as bacterial cells, is similar to that used for detecting a protein analyte that has two binding sites for two or more MAbs. In this case, the binding sites may comprise the same protein embedded in the cell surface.
- the binding pair reagent spans the distance between individual protein units.
- the cells that may be detected according to this method include bacterial, animal, plant, insect and fungal cells.
- the animal cell is a human cell, which may be diseased or healthy.
- the method may be used to detect circulating cancer cells by employing binding pair comprising antibodies to cell surface tumor antigens. Analyte Detection Using Binding Pairs Having 3'-5' Conjugation of Overlap DNA Markers
- binding pair reagents comprising one binding pair member (5' conjugate) comprising a first nucleic acid of an overlapping pair conjugated to the first specificity molecule (e.g. first MAb of a sandwich pair) at the 5' end and the other binding pair member (3'conjugate) comprising the complementary second nucleic acid strand of an overlapping pair conjugated to a second specificity molecule (e.g. second Mab) via the 3' end as illustrated in FIG. 6.
- the 3' end of the 5' conjugate hybridizes to an internal sequence of the 3' conjugate, producing a binding pair where extension of the 3' end is directed away from the center of mass of the formed complex and towards a freely rotating terminus (the 5' end of the 3' complex).
- This orientation improves the efficiency of chain extension when compared to 3 '-3' overlap binding pairs thereby allowing for greater steric access of polymerase and the dissipation of torsional stress during initial polymerization.
- the DNA/RNA labels are complementary on the 3' ends for the entire stretch of RNA sequence and for short stretch of DNA sequence.
- the RNA sequence is at least about 20 bases long. In other embodiments the RNA sequence is at least about 30, 40, 50 or 60 bases long. According to one aspect of the invention, the RNA sequence is 26 bases long.
- the DNA-DNA duplex region is short, typically comprising less than 15 base pairs, frequently less than about 12 base pairs, and preferably less than about 10 base pairs in length. In various aspects of the invention, the DNA-DNA duplex is 12, 11, 10, 9, 8 or 7 base pairs in length.
- a nucleic acid duplex region of the analyte-binding pair complex is stable at elevated temperatures, thereby ensuring the formation of a complex containing binding members locked into a specific orientation and distance from each other in the binding pair complex.
- the RNA component of both the complexed and excess binding pair can be digested by the addition of RNase, leaving a short, relatively unstable binding pair joined by the residual DNA/DNA duplex.
- the RNase is RNase H. Following RNase treatment, the unstable binding pair dissociates into its respective binding members upon raising the temperature. Only the binding members fixed into position on the complex rapidly reassociate upon reducing the temperature.
- binding members with nucleic acid components of shortened 3' DNA overlap reduces the requirement for dilution of the complex formation reaction and, therefore, improves sensitivity further.
- Nucleic acid strands comprising both deoxyribose and ribose nucleotide monomer, and having a 5' amino function may be synthesized by standard phosphoramidite chemistry. Alternatively, a short 3' DNA/DNA overlap may be filled in enzymatically with RNA polymerase. Sensitive Homogeneous Assay for Analytes
- a homogeneous assay has been devised for analytes such as protein antigens and microorganism that approaches the sensitivity achieved for nucleic acid detection.
- a first specificity molecule e.g. a first MAb of a sandwich pair or a first portion of polyclonal antibody
- the RNA portion of the DNA/RNA oligo comprises 30 bases at the 3' end.
- a unique primer site is located at the 5' DNA terminus of the DNA/RNA oligo.
- the second specificity molecule (e.g., second MAb or a second portion of polyclonal antibody) is conjugated to the 3' end of a DNA oligonucleotide optionally synthesized with one or more 3' phosphoramidite spacers to increase overall length.
- the DNA oligo is at least 60 bases long.
- a unique primer binding site is located at the 5' end of the DNA oligo.
- the oligonucleotides form a duplex of relatively long RNA-DNA hybrid, typically 20-40 base pairs in length, followed by a short stretch of DNA-DNA hybrid, of approximately 7-15 base pairs. (FIG. 7).
- the binding pairs of this embodiment can be extended by polymerase in only one direction from the exposed 3' DNA terminus (FIG. 7). Steric and torsional constraints imposed by 3' overlaps are eliminated, as described above. Enzymatic digestion of the binding pair results in a short, DNA-DNA duplex, which is relatively unstable and thereby minimizes reformation of binding pair from free binding members in the bulk solution.
- Two 60-base single-strand DNA sequences [SEQ ID NOS. 1 and 3] were synthesized to contain a single primary amine group attached to the 5' end via a 12- carbon spacer arm (Glen Research, Sterling, VA). The sequences were complimentary to each other for the last 15 bases on the 3' terminus. Both were treated identically.
- the 5' amino-DNA (350 ⁇ g) was dissolved in 25 ⁇ l 0.05 M sodium phosphate buffer, pH 7.0.
- Disuccinimidyl suberate was dissolved in dry DMSO at a concentration of 10 mg/ml, and a 50 ⁇ l aliquot was added immediately to the DNA solution and mixed by gentle re- pipetting.
- reaction mixture was injected onto a 0.5 x 20 cm G-25 (fine) column plumbed to a Pharmacia FPLC system.
- the column was equilibrated in 5 mM citric acid, adjusted to pH 5.4 with 0.1 M sodium hydroxide, and developed at a flow rate of 0.5 ml/min at room temperature.
- the first eluted peak was collected directly into an Microcon ® YM-IO centrifugal ultra filtration (Millipore, Billerica, Mass.) unit embedded in ice.
- the suberate-activated template DNA was concentrated at 4° C and 5000 x g for 30 minutes, to 200-300 ⁇ l. The DNA concentration was quickly assessed by absorbance at 260 nm.
- the activity of the lyophilized, activated DNA was assessed by conjugation to a small amine-containing molecule.
- Glutathione at 50 mg/ml in 0.1 M sodium phosphate, pH 8.0 was reacted with a stoichiometric amount of N-ethyl maleimide (NEM).
- NEM N-ethyl maleimide
- the pH was re-adjusted to 8.0 with sodium hydroxide.
- One A 260 unit of suberate-activated DNA was dissolved in 10 ⁇ l of NEM-treated glutathione, then diluted 25-fold with water to 4.0 A 26 o units/ml.
- sandwich-paired MAbs directed against different epitopes on PSA, were obtained from Bios Pacific, Inc. (Emeryville, CA) and each was conjugated to one of the activated overlap DNA sequences.
- MAb 1 mg/ml was buffer exchanged and concentrated into 0.1 M sodium phosphate, 0.15 M sodium chloride, pH 8.25 by two passes through a Microcon ® 50 centrifugal ultra filtration unit (Millipore, Billerica, Mass.) at 10,000 x g in a micro centrifuge at room temperature.
- the final volume of 25-50 ⁇ l was collected by centrifugation into a micro centrifuge tube and added to a lyophilized pellet of activated DNA (prepared as described above in Example 1).
- the conjugation reaction was allowed to proceed for several hours at room temperature.
- the degree of antibody labeling with DNA was assessed by SDS and/or native protein gel electrophoresis.
- the incorporation of one or more strands of DNA into an antibody structure increased both the molecular weight and electronegativity.
- Antibody labeled with DNA appeared as distinct, slower migrating bands when electrophoresed on 4% polyacrylamide-SDS gels, with the rate of electrophoretic migration corresponding to the degree of DNA incorporation.
- the same conjugated molecules migrated faster than the corresponding underivatized antibody on 4-12% native protein gel electrophoresis. The separation was more dramatic, but less defined for higher order conjugates on native gels.
- the extent of DNA conjugation varied from 50 to 100%, depending on the relative reactivity and exposure of lysine amine groups in each antibody structure.
- the conjugates were found to have an average labeling density of 2.5.
- the reaction mixture containing antibody-DNA conjugates was added to a second lyophilized pellet of activated DNA to increase the extent of antibody labeling with DNA.
- the MAb-DNA conjugates were individually concentrated by centrifugal ultrafiltration and assayed for protein using a BCA Protein Assay (Pierce Biotechnology, Inc., Rockford, IL). Equal amounts of both conjugates were combined to form the binding pair, which hybridize to form a single molecule. Sodium azide was added to a final concentration of 0.1% and the binding pair reagent was stored at 4° C.
- BCA Protein Assay Pierce Biotechnology, Inc., Rockford, IL
- the binding pair of Example 2 was diluted to 20 pM and combined with an equal volume of a dilution series of antigen containing 0 to 1 ng/ml PSA.
- the antigen-binding pair mixture was incubated for two hours at 37° C. The incubation allowed formation of binding complexes comprising one antigen molecule and one binding pair molecule, with the antibody regions of each member of the binding pair bound to a distinct epitope on the PSA antigen. All salts were intentionally omitted to diminish hybridization of the overlap oligonucleotide labels in the reaction solution.
- the complementary DNA regions of complexed binding pair members re-anneal rapidly to reform the overlap hybrid of the binding pair reagent due to their close proximity and orientation.
- the DNA regions of complexed binding pairs can be extended by Taq polymerase, thereby forming a template for PCR.
- Uncomplexed, free binding pair members do not re-anneal to a significant extent prior to or during the amplification reaction because they are too dilute.
- the reaction solution 50 ⁇ l
- the temperature was ramped from 50° C to 85° C over 3 minutes and then held at 95° C for 2 minutes.
- Example 4 Homogeneous Assay for E. coli 0157
- E. coli cells were obtained from ATCC (Manassas, VA) and grown in liquid culture. Cell concentration was determined by the appearance of colonies on agar media grown overnight at 37° C.
- Polyclonal antibodies against E. coli 0157 were obtained from KPL, Inc. (Gaithersburg, Maryland) and conjugated to nucleic acid marker oligos having complementary overlapping sequences, according to the procedures described in Examples 1 and 2. Aliquots of the same polyclonal antibody preparation were labeled individually, each with one of the two overlap oligonucleotides, purified and combined stoichiometrically to produce a binding pair reagent.
- Bacterial cells were suspended in neutral buffer (10 mM Tris, pH 7.5) to stop active growth and treated with an equal volume of 20 to 200 pM binding pair reagent for two hours at room temperature.
- the binding of one antibody member of the binding pair to a cell surface protein enhances the binding of the second member, forming a molecular bridge joining two adjacent binding sites on a cell.
- the cell suspension was then diluted 1:100 in 10 mM Tris, pH 7.5, and warmed to 45°-50° C to dissociate excess binding pair reagent to independent binding pair members.
- An equal volume of 2X PCR mix containing 400 ⁇ M primers was added and the mixture incubated for 3 minutes at room temperature.
- the reaction (50 ⁇ l) was transferred to the well of a microtiter plate, sealed and subjected to real-time PCR under the conditions described above in Example 3. Prior to PCR, a sample was also removed for determination of bacterial cell number by plating onto solid media. Following 45 amplification cycles, a lower limit of 10-50 intact cells were detected using this method.
- binding pair reagents comprising one binding pair member composed of one DNA strand of an overlapping pair conjugated to one MAb of a sandwich pair at the 5' end as described in Example 1 and the second DNA strand conjugated to the other MAb via the 3' end.
- the 3' end of the 5' conjugate hybridizes to an internal sequence of the 3' conjugate, producing a binding pair where extension of the free 3' end occurs away from the center of mass of a formed complex and towards a freely rotating terminus.
- Such an orientation may improve the efficiency of chain extension over previously described 3 '-3' overlap binding pairs by allowing for greater steric access of polymerase and the dissipation of torsional stress during initial polymerization.
- the binding members of this example are formed and purified as described in Examples 1 and 2, using monoclonal antibody sandwich pair obtained from Roche Diagnostics, Indianapolis, IN.
- the 5' DNA labels [SEQ ID NOS. 6 and 7] were comprised of 60 bases having the 3' terminus complimentary for 9 bases.
- SEQ ID NO. 6 was complimentary with an internal 9 base sequence of a 3' DNA label of 75 bases [SEQ ED NO 8].
- the 3' DNA label may also be synthesized to contain one or several Spacer Phosphoramidites (Glenn Research) on the 3 'end to compensate for the lost length of the binding pair crated by the internalized hybridization.
- SEQ ID NO. 6 was arbitrarily conjugated to monoclonial antibody designated, MAK ⁇ TNT>M-7.
- SEQ ID NOS. 7 and 8 were conjugated to monoclonial antibody designated, MAK ⁇ TN-T>M- 11-7. The conjugation chemistry and purification remained the same.
- the binding pair comprising a stable internal overlap hybrid segment is diluted, reacted with Troponin-T and amplified in the presence of downstream primers (SEQ ID NOS. 9 and 10) as described for Example 3.
- the free and complexed binding pair has similar melting and re-association properties as that of a 3 '-3' overlap binding pair of the same sequence.
- the outward orientation of initial polymerization of a 3'-5- overlap improves efficiency of template sequence generation in those instances where the sandwich complex bulk may hinder the progression of the Taq polymerase. Amplification of a greater number of initial template molecules allows faster signal threshold in real-time PCR and greater assay sensitivity.
- the homogeneous assay for Troponin-T using a 3 '-3' overlap binding pair reagent exhibited detection sensitivity similar to that of PSA (example 3).
- the corresponding homogeneous assay for Troponin- T using the 3 '-5' overlap orientation yielded a 2-4 fold improvement.
- Example 6 Homogeneous Assay for Streptavidin using 3'-3' and 3'-5' Biotin - Labeled Overlap DNA.
- Streptavidin serves as the "antigen" in this model system, having a molecular diameter of approximately 60 angstroms.
- the terminal amino functions of SEQ ID NOS. 6, 7 and 8 were labeled with sulfosuccinimidyl 2-(biotinamido) ethyl-l,3'dithiopropionate (Pierce Biotechnology, Inc., Rockford, IL) and purified by FPLC gel filtration chromatography.
- Each DNA label is over 200 angstroms in length when folly extended, and guarantees ability to span the distance required to achieve overlap hybridization.
- Streptavidin (Prozyme, San Leandro, CA) was dissolved in 10 mM Tris containing 0.05% tween-20 and 0.05% sodium azide, pH 8.0 and allowed to form a complex with both 3 '-3' and 3 '-5' overlap DNA labeled with biotin, present at a final concentration of 10-100 pM. The complex was detected by treatnent with the addition 2X PCR mix for 15 minutes, warmed to 37 0 C, and subjected to real-time PCR cycling in the presence of downstream primers (SEQ ED NOS. 9 and 10). Streptavidin could be detected at an input of approximately 500 molecules (50 attograms) when assayed in the 3'-5' overlap configuration and 2000 molecules in the 3 '-3' orientation.
- the binding members of this method can be formed and purified as described in Examples 1 and 2, except that one of the nucleic acids is coupled via its 3' end.
- the 5' DNA label is a 60 base oligonucleotide with a 3' terminus complimentary to 15 bases of an internal sequence of the 3' DNA label.
- the 3' DNA label is synthesized to contain spacer phosphoramidites (Glenn Research, Sterling, VA) on the 3' end to compensate for the lost length of the binding pair created by the internalized hybridization.
- the conjugation chemistry and purification are performed as described above.
- the binding pair comprising a stable internal overlap hybrid segment is diluted, reacted with PSA and amplified as described in Example 3.
- the free and complexed binding pairs have similar melting and re-association properties as 3 '-3' overlapped binding pairs, which are dictated by the thermodynamic properties of DNA base-pairing.
- the outward orientation of initial polymerization improves efficiency of template sequence generation. Amplification of a greater number of initial template molecules allows faster signal threshold in real-time PCR and greater assay sensitivity.
- Example 8 Homogeneous Assay of PSA Employing Enzymatic Digestion
- binding members comprising antibody conjugated to single-strand nucleic acids with deoxyribose bases at the 5' end and ribose bases at the 3' end.
- Nucleic acid strands composed of both deoxyribose and ribose nucleotide monomer containing a 5' amino function are synthesized by standard phosphoramidite chemistry.
- a short 3' DNA/DNA overlap may be filled in enzymatically with RNA polymerase.
- Overlapping nucleic acid labels made up of both DNA and RNA are synthesized starting with RNA phosphoramidite for the first 26 bases.
- the remaining 59 bases are synthesized with DNA phosphoramidite, terminating in a (C 12 ) amino function.
- the first 25 bases from the 5' ends represent unique primer sequences.
- the next 26 DNA bases are complimentary to the 26 RNA bases of the overlapping strand.
- the next 9 DNA bases are complimentary to the next 9 DNA bases of the overlapping strand, thus the DNA/RNA labels are complementary on the 3' ends for the entire stretch of RNA sequence and for about 9 bases of DNA sequence.
- the binding pair consists of two binding members attached via a relatively lengthy stretch of DNA/RNA duplex and a relatively short stretch of DNA/DNA duplex. When conjugated to sandwich MAbs against PSA, as described in Examples 2 & 3, and combined stoichiometrically, the resulting binding pair contains a 110 base nucleotide bridge of which 60 internal bases were paired.
- This reagent is used as in Example 3 to detect PSA.
- the binding pair forms a complex with analyte by recognition of two PSA epitopes by the two antibody members of the pair.
- the nucleic acid duplex region of the antigen-binding pair complex particularly the DNA/RNA heteroduplex region, is stable at elevated temperatures and ensures the formation of a complex containing binding members locked into a specific orientation and distance from each other in the binding pair complex.
- the reaction is diluted only 10-fold and treated with RNAse H (Promega Corp., Madison, WI) for 30 min.
- the resulting binding pair is thermally dissociated by warming to 45° C for 3 minutes, then cooled to room temperature.
- the remaining short DNA/DNA overlap reassociate rapidly, if only transiently, in the immune complex. Only the binding members fixed into position on the complex rapidly reassociate upon reducing the temperature. The dissociated binding members are unable to reform binding pairs due to their low concentration and the limited stability of the short DNA/DNA overlap.
- binding members with DNA labels of shortened 3' overlap reduces the requirement for dilution of the complex formation reaction and, therefore, improves sensitivity of the assay as compared to the assay of Example 3, above.
- PCR mix and primers are added, and real-time PCR amplification is performed as described above.
- Example 9 Homogeneous Assay for PSA Using Enzymatic Digestion and 3'-5' Conjugation.
- a homogeneous assay can be devised for protein antigens and microorganism that approaches the sensitivity achieved for nucleic acid detection.
- the first MAb of a sandwich pair is conjugated to the 5' end of a chimeric DNA/RNA oligonucleotide. It will be appreciated that a first portion of polyclonal antibody can be used in place of the Mab.
- the RNA portion of the DNA/RNA oligo comprises 30 bases at the 3' end.
- a unique primer site is located at the 5' DNA terminus of the DNA/RNA oligo.
- the second Mab (which could be substituted by a second portion of polyclonal antibody) is conjugated to the 3' end of a 64-mer DNA oligonucleotide which is synthesized with several 3' phosphoramidite spacers to increase overall length.
- the 5' terminal 25 bases from the 5' end of the 64-mer represent a unique primer site.
- the next 30 bases are complimentary to the 30 RNA bases of the DNA/RNA oligo.
- the last 9 bases are complimentary to the DNA segment adjacent to the RNA segment (FIG. 7).
- both binding members produces a binding pair with an internal 9 base pair DNA/DNA hybrid adjacent to a 30 base pair DNA/DNA hybrid, as illustrated in Figure 7.
- the binding pair could be extended by polymerase in only one direction from the exposed 3' DNA terminus (see Fig. 7).
- Steric and torsional constraints imposed by 3' overlaps are eliminated, as in Example 5.
- Enzymatic digestion of the binding pair results in a short 9 base pair overlap, which is relatively unstable and minimizes reformation of the binding pair in the bulk solution.
- PCR amplification is carried out as described above.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Pathology (AREA)
- Urology & Nephrology (AREA)
- Medicinal Chemistry (AREA)
- Hematology (AREA)
- Crystallography & Structural Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Computational Biology (AREA)
- Food Science & Technology (AREA)
- Plant Pathology (AREA)
- Gynecology & Obstetrics (AREA)
- Pregnancy & Childbirth (AREA)
- Reproductive Health (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analyzing Materials By The Use Of Fluid Adsorption Or Reactions (AREA)
Abstract
Description
Claims
Priority Applications (11)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2007539365A JP5188808B2 (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
EP05858303.0A EP1842226B2 (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
AU2005333156A AU2005333156B2 (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
ES05858303.0T ES2352041T5 (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
CA002585781A CA2585781A1 (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
AT05858303T ATE480643T1 (en) | 2004-11-03 | 2005-11-03 | HOMOGENEOUS DETECTION OF ANALYTES |
CN2005800379854A CN101137758B (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
US11/718,379 US8338579B2 (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
DK05858303.0T DK1842226T3 (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
DE602005023529T DE602005023529D1 (en) | 2004-11-03 | 2005-11-03 | HOMOGENEOUS DETECTION OF ANALYTES |
US13/117,816 US9234890B2 (en) | 2004-11-03 | 2011-05-27 | Homogeneous analyte detection |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US62495004P | 2004-11-03 | 2004-11-03 | |
US60/624,950 | 2004-11-03 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/718,379 A-371-Of-International US8338579B2 (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
US13/117,816 Division US9234890B2 (en) | 2004-11-03 | 2011-05-27 | Homogeneous analyte detection |
Publications (3)
Publication Number | Publication Date |
---|---|
WO2006137932A2 true WO2006137932A2 (en) | 2006-12-28 |
WO2006137932A9 WO2006137932A9 (en) | 2007-02-15 |
WO2006137932A3 WO2006137932A3 (en) | 2007-09-20 |
Family
ID=37570907
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2005/040133 WO2006137932A2 (en) | 2004-11-03 | 2005-11-03 | Homogeneous analyte detection |
Country Status (13)
Country | Link |
---|---|
US (2) | US8338579B2 (en) |
EP (1) | EP1842226B2 (en) |
JP (2) | JP5188808B2 (en) |
KR (1) | KR20070105967A (en) |
CN (2) | CN101137758B (en) |
AT (1) | ATE480643T1 (en) |
AU (1) | AU2005333156B2 (en) |
CA (1) | CA2585781A1 (en) |
DE (1) | DE602005023529D1 (en) |
DK (1) | DK1842226T3 (en) |
ES (1) | ES2352041T5 (en) |
PT (1) | PT1842226E (en) |
WO (1) | WO2006137932A2 (en) |
Cited By (62)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009105264A1 (en) * | 2008-02-21 | 2009-08-27 | Iris International Inc. | Method for early determination of recurrence after therapy for prostate cancer |
WO2012085113A1 (en) * | 2010-12-23 | 2012-06-28 | Roche Diagnostics Gmbh | Binding agent |
WO2012085111A1 (en) | 2010-12-23 | 2012-06-28 | F. Hoffmann-La Roche Ag | Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery |
WO2012104261A1 (en) | 2011-01-31 | 2012-08-09 | Olink Ab | Exonuclease enabled proximity extension assays |
US8513032B2 (en) | 2004-11-03 | 2013-08-20 | Iris International, Inc. | Microbubbles for affinity separation |
WO2014001326A1 (en) | 2012-06-27 | 2014-01-03 | F. Hoffmann-La Roche Ag | Method for the selection and production of tailor-made, selective and multi-specific therapeutic molecules comprising at least two different targeting entities and uses thereof |
WO2013113699A3 (en) * | 2012-01-30 | 2014-03-20 | Olink Ab | Hyperthermophilic polymerase enabled proximity extension assay |
US9234890B2 (en) | 2004-11-03 | 2016-01-12 | Iris International, Inc. | Homogeneous analyte detection |
WO2016080838A1 (en) * | 2014-11-21 | 2016-05-26 | Piculet Biosciences Technologies B.V. | Self-assembled bivalent ligand complex (sablc) libraries and methods for screening such libraries |
CN106124781A (en) * | 2016-07-07 | 2016-11-16 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of method based on aptamers sandwich fluorescent quenching technology for detection demethyl testosterone |
US9688758B2 (en) | 2012-02-10 | 2017-06-27 | Genentech, Inc. | Single-chain antibodies and other heteromultimers |
US9761637B2 (en) | 2010-06-16 | 2017-09-12 | Sony Corporation | Display device |
US9768666B2 (en) | 2011-09-08 | 2017-09-19 | Siemens Aktiengesellschaft | External cooling tube arrangement for a stator of an electric motor |
US9862779B2 (en) | 2012-09-14 | 2018-01-09 | Hoffmann-La Roche Inc. | Method for the production and selection of molecules comprising at least two different entities and uses thereof |
US9994646B2 (en) | 2009-09-16 | 2018-06-12 | Genentech, Inc. | Coiled coil and/or tether containing protein complexes and uses thereof |
US10106612B2 (en) | 2012-06-27 | 2018-10-23 | Hoffmann-La Roche Inc. | Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof |
US10106600B2 (en) | 2010-03-26 | 2018-10-23 | Roche Glycart Ag | Bispecific antibodies |
US10131958B1 (en) | 2013-08-28 | 2018-11-20 | Cellular Research, Inc. | Massively parallel single cell analysis |
WO2018226293A1 (en) * | 2017-06-05 | 2018-12-13 | Becton, Dickinson And Company | Sample indexing for single cells |
US10202646B2 (en) | 2009-12-15 | 2019-02-12 | Becton, Dickinson And Company | Digital counting of individual molecules by stochastic attachment of diverse labels |
US10338066B2 (en) | 2016-09-26 | 2019-07-02 | Cellular Research, Inc. | Measurement of protein expression using reagents with barcoded oligonucleotide sequences |
US10633457B2 (en) | 2014-12-03 | 2020-04-28 | Hoffmann-La Roche Inc. | Multispecific antibodies |
US10640763B2 (en) | 2016-05-31 | 2020-05-05 | Cellular Research, Inc. | Molecular indexing of internal sequences |
GB202004469D0 (en) | 2020-03-27 | 2020-05-13 | Olink Proteomics Ab | Controls for proximity detection assays |
GB202004472D0 (en) | 2020-03-27 | 2020-05-13 | Olink Proteomics Ab | Method for detecting analytes of varying abundance |
US10781473B2 (en) | 2015-10-21 | 2020-09-22 | Olink Proteomics Ab | Method for generating proximity probes |
GB202018503D0 (en) | 2020-11-25 | 2021-01-06 | Olink Proteomics Ab | Analyte detection method employing concatamers |
US10941396B2 (en) | 2012-02-27 | 2021-03-09 | Becton, Dickinson And Company | Compositions and kits for molecular counting |
EP3580356B1 (en) * | 2017-02-09 | 2021-03-31 | Atlas Antibodies AB | Method for determining levels of interactions between biomolecules |
US10982263B2 (en) | 2014-06-23 | 2021-04-20 | The Board Of Trustees Of The Leland Stanford Junior University | On-slide staining by primer extension |
US10982007B2 (en) | 2010-12-23 | 2021-04-20 | Roche Diagnostics Operations, Inc. | Detection of a posttranslationally modified polypeptide by a bivalent binding agent |
WO2021191448A1 (en) | 2020-03-27 | 2021-09-30 | Olink Proteomics Ab | Method for detecting analytes |
US11168350B2 (en) | 2016-07-27 | 2021-11-09 | The Board Of Trustees Of The Leland Stanford Junior University | Highly-multiplexed fluorescent imaging |
USRE48913E1 (en) | 2015-02-27 | 2022-02-01 | Becton, Dickinson And Company | Spatially addressable molecular barcoding |
US11300703B2 (en) | 2015-03-20 | 2022-04-12 | Rapiscan Systems, Inc. | Hand-held portable backscatter inspection system |
US20220119552A1 (en) * | 2020-10-15 | 2022-04-21 | City Of Hope | Complementary rna linked bispecific t-cell engaging antibodies |
US11319583B2 (en) | 2017-02-01 | 2022-05-03 | Becton, Dickinson And Company | Selective amplification using blocking oligonucleotides |
US11332776B2 (en) | 2015-09-11 | 2022-05-17 | Becton, Dickinson And Company | Methods and compositions for library normalization |
WO2022101414A1 (en) | 2020-11-12 | 2022-05-19 | Oslo Universitetssykehus Hf | Method of determining dlbcl prognosis |
US11340361B1 (en) | 2020-11-23 | 2022-05-24 | American Science And Engineering, Inc. | Wireless transmission detector panel for an X-ray scanner |
US11365409B2 (en) | 2018-05-03 | 2022-06-21 | Becton, Dickinson And Company | Molecular barcoding on opposite transcript ends |
US11390914B2 (en) | 2015-04-23 | 2022-07-19 | Becton, Dickinson And Company | Methods and compositions for whole transcriptome amplification |
US11421022B2 (en) | 2012-06-27 | 2022-08-23 | Hoffmann-La Roche Inc. | Method for making antibody Fc-region conjugates comprising at least one binding entity that specifically binds to a target and uses thereof |
US11492660B2 (en) | 2018-12-13 | 2022-11-08 | Becton, Dickinson And Company | Selective extension in single cell whole transcriptome analysis |
US11525157B2 (en) | 2016-05-31 | 2022-12-13 | Becton, Dickinson And Company | Error correction in amplification of samples |
US11525930B2 (en) | 2018-06-20 | 2022-12-13 | American Science And Engineering, Inc. | Wavelength-shifting sheet-coupled scintillation detectors |
US11535882B2 (en) | 2015-03-30 | 2022-12-27 | Becton, Dickinson And Company | Methods and compositions for combinatorial barcoding |
US11579327B2 (en) | 2012-02-14 | 2023-02-14 | American Science And Engineering, Inc. | Handheld backscatter imaging systems with primary and secondary detector arrays |
US11639517B2 (en) | 2018-10-01 | 2023-05-02 | Becton, Dickinson And Company | Determining 5′ transcript sequences |
US11649497B2 (en) | 2020-01-13 | 2023-05-16 | Becton, Dickinson And Company | Methods and compositions for quantitation of proteins and RNA |
US11661625B2 (en) | 2020-05-14 | 2023-05-30 | Becton, Dickinson And Company | Primers for immune repertoire profiling |
US11661631B2 (en) | 2019-01-23 | 2023-05-30 | Becton, Dickinson And Company | Oligonucleotides associated with antibodies |
US11739443B2 (en) | 2020-11-20 | 2023-08-29 | Becton, Dickinson And Company | Profiling of highly expressed and lowly expressed proteins |
US11773436B2 (en) | 2019-11-08 | 2023-10-03 | Becton, Dickinson And Company | Using random priming to obtain full-length V(D)J information for immune repertoire sequencing |
US11773441B2 (en) | 2018-05-03 | 2023-10-03 | Becton, Dickinson And Company | High throughput multiomics sample analysis |
US11932849B2 (en) | 2018-11-08 | 2024-03-19 | Becton, Dickinson And Company | Whole transcriptome analysis of single cells using random priming |
US11932901B2 (en) | 2020-07-13 | 2024-03-19 | Becton, Dickinson And Company | Target enrichment using nucleic acid probes for scRNAseq |
US11939622B2 (en) | 2019-07-22 | 2024-03-26 | Becton, Dickinson And Company | Single cell chromatin immunoprecipitation sequencing assay |
US11965208B2 (en) | 2019-04-19 | 2024-04-23 | Becton, Dickinson And Company | Methods of associating phenotypical data and single cell sequencing data |
WO2024100258A1 (en) | 2022-11-11 | 2024-05-16 | Olink Proteomics Ab | Library of proximity probes and method of use |
EP4382534A1 (en) | 2022-12-06 | 2024-06-12 | Biomérieux | Synthetic peptide for use in an immunomolecular assay |
US12071617B2 (en) | 2019-02-14 | 2024-08-27 | Becton, Dickinson And Company | Hybrid targeted and whole transcriptome amplification |
Families Citing this family (48)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ES2463420T3 (en) * | 2002-11-01 | 2014-05-27 | Iris International, Inc. | Immuno-PCR sandwich sandwich |
EP2235536A4 (en) | 2007-12-20 | 2011-05-04 | Lab Corp America Holdings | Her-2 diagnostic methods |
WO2010011901A1 (en) * | 2008-07-24 | 2010-01-28 | Innovative Biosensors, Inc. | Detection of rna-dna duplex molecules |
US8349574B2 (en) | 2009-01-15 | 2013-01-08 | Laboratory Corporation Of America Holdings | Methods of determining patient response by measurement of Her-3 |
CA2764386C (en) | 2008-12-01 | 2018-05-01 | Laboratory Corporation Of America Holdings | P95-her2 antibodies and uses thereof |
SG172984A1 (en) * | 2009-01-15 | 2011-08-29 | Lab Corp America Holdings | Methods of determining patient response by measurement of her-2 expression |
KR101029343B1 (en) * | 2009-07-30 | 2011-04-13 | 한국과학기술연구원 | Immunoassay-based Antigen Detecting Kit and Method |
CA2817455C (en) * | 2010-12-23 | 2019-04-16 | F. Hoffmann-La Roche Ag | Detection of a polypeptide dimer by a bivalent binding agent |
EP2882869A4 (en) | 2012-08-07 | 2016-04-20 | Jackson H M Found Military Med | Prostate cancer gene expression profiles |
EP2971121A4 (en) | 2013-03-15 | 2016-11-30 | Fluidigm Corp | Simultaneous detection of target protein and target nucleic acids in a single cell |
US9593364B2 (en) * | 2013-05-21 | 2017-03-14 | Src, Inc. | Detecting a target molecule in a sample using a dual-antibody quantitative fluorescence-based detection method |
EP3041957A4 (en) * | 2013-09-04 | 2017-03-29 | Fluidigm Corporation | Proximity assays for detecting nucleic acids and proteins in a single cell |
US10711311B2 (en) | 2013-12-30 | 2020-07-14 | The Henry M. Jackson Foundation For The Advancement Of Military Medicine, Inc. | Genomic rearrangements associated with prostate cancer and methods of using the same |
CN103728452B (en) * | 2014-01-23 | 2015-06-24 | 湖南大学 | Colloidal gold immunoassay kit for detecting aflatoxin B1 and preparation method of kit |
CN103728451B (en) * | 2014-01-23 | 2015-06-24 | 湖南大学 | Colloidal gold immunoassay kit for detecting ochracin A and preparation method of kit |
US11291931B2 (en) | 2014-12-15 | 2022-04-05 | Akadeum Life Sciences, Inc. | Method and system for buoyant separation |
JP6822973B2 (en) * | 2015-04-17 | 2021-01-27 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | A method for detecting agglomeration and a composition used in carrying out the method. |
CN108780081B (en) | 2015-08-10 | 2021-04-09 | Essenlix公司 | Simplified-procedure, small-sample, rapid, easy-to-use bio/chemical analysis apparatus and method |
SG10202104563UA (en) | 2015-09-14 | 2021-06-29 | Essenlix Corp | Device and system for analyzing a sample, particularly blood, as well as methods of using the same |
KR20190057446A (en) | 2015-09-14 | 2019-05-28 | 에센릭스 코프. | Device and system for collecting and analyzing vapor condensate, particularly exhaled breath condensate, as well as method of using the same |
US10301677B2 (en) | 2016-05-25 | 2019-05-28 | Cellular Research, Inc. | Normalization of nucleic acid libraries |
DE102016221875B4 (en) * | 2016-11-08 | 2018-06-28 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Stationary phase for detecting a particular analyte in a mixture, uses thereof, and method for detecting a particular analyte in a mixture |
US20180168196A1 (en) | 2016-12-15 | 2018-06-21 | Nestec Sa | Compositions and methods that modulate bacteria in a companion animal |
WO2018119318A1 (en) | 2016-12-21 | 2018-06-28 | Essenlix Corporation | Devices and methods for authenticating a sample and use of the same |
EP3579981A4 (en) | 2017-02-07 | 2021-03-31 | Essenlix Corporation | Compressed open flow assay and use |
WO2018148470A1 (en) | 2017-02-08 | 2018-08-16 | Essenlix Corporation | Sample collection and handling for delayed analysis |
CN111936837B (en) | 2017-02-08 | 2024-06-07 | 上海宜晟生物科技有限公司 | QMAX assay and use |
WO2018148469A1 (en) | 2017-02-08 | 2018-08-16 | Essenlix Corp. | Bio/chemical material extraction and assay |
WO2018148609A2 (en) | 2017-02-09 | 2018-08-16 | Essenlix Corporation | Colorimetric assays |
US11883824B2 (en) | 2017-02-09 | 2024-01-30 | Essenlix Corporation | Assay using different spacing heights |
CA3053132A1 (en) | 2017-02-09 | 2018-08-16 | Essenlix Corp. | Assay with amplification |
CA3053301A1 (en) | 2017-02-16 | 2018-08-23 | Essenlix Corporation | Assay with textured surface |
CN112689758A (en) | 2017-08-01 | 2021-04-20 | Essenlix公司 | Device and method for examining the effect of a drug on microorganisms |
CN111492222A (en) | 2017-08-01 | 2020-08-04 | Essenlix公司 | Sample collection, retention and assay |
US11280706B2 (en) | 2017-08-01 | 2022-03-22 | Essenlix Corporation | Dilution calibration |
WO2019075415A1 (en) | 2017-10-13 | 2019-04-18 | Essenlix Corporation | Devices and methods for authenticating a medical test and use of the same |
US11237113B2 (en) | 2017-10-26 | 2022-02-01 | Essenlix Corporation | Rapid pH measurement |
US11609224B2 (en) | 2017-10-26 | 2023-03-21 | Essenlix Corporation | Devices and methods for white blood cell analyses |
US10807095B2 (en) | 2017-10-26 | 2020-10-20 | Essenlix Corporation | Making and tracking assay card |
US11648551B2 (en) | 2017-12-12 | 2023-05-16 | Essenlix Corporation | Sample manipulation and assay with rapid temperature change |
WO2019118936A2 (en) | 2017-12-14 | 2019-06-20 | Essenlix Corporation | Devices, systems, and methods for monitoring hair |
WO2019140334A1 (en) | 2018-01-11 | 2019-07-18 | Essenlix Corporation | Homogeneous assay (ii) |
WO2019165129A1 (en) * | 2018-02-22 | 2019-08-29 | The Board Of Trustees Of The Leland Stanford Junior University | Methods for diagnosing and for determining severity of an autism spectrum disorder |
WO2020014273A1 (en) | 2018-07-09 | 2020-01-16 | Akadeum Life Sciences, Inc. | System and method for buoyant particle processing |
US11885952B2 (en) | 2018-07-30 | 2024-01-30 | Essenlix Corporation | Optics, device, and system for assaying and imaging |
JP2022518515A (en) * | 2019-01-25 | 2022-03-15 | プソマーゲン, インコーポレイテッド | Detection and treatment of antibody-DNA complex and HPV |
CN112730840B (en) * | 2021-01-29 | 2023-10-17 | 章毅 | Method for identifying CD44 and CD24 molecular phenotypes |
WO2023028329A1 (en) | 2021-08-26 | 2023-03-02 | Akadeum Life Sciences, Inc. | Method and system for buoyant separation |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
US5635602A (en) | 1993-08-13 | 1997-06-03 | The Regents Of The University Of California | Design and synthesis of bispecific DNA-antibody conjugates |
US20050026161A1 (en) | 2002-11-01 | 2005-02-03 | Edward Jablonski | Displacement sandwich immuno-PCR |
Family Cites Families (66)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB1582956A (en) | 1976-07-30 | 1981-01-21 | Ici Ltd | Composite magnetic particles |
US4376110A (en) | 1980-08-04 | 1983-03-08 | Hybritech, Incorporated | Immunometric assays using monoclonal antibodies |
DE3100535A1 (en) | 1981-01-10 | 1982-08-12 | Dr. Karl Thomae Gmbh, 7950 Biberach | "NEW CARBONIC ACID DERIVATIVES, THEIR PRODUCTION AND THEIR USE AS MEDICINAL PRODUCTS" |
FR2571498B1 (en) | 1984-10-04 | 1988-04-08 | Immunotech Sa | METHOD FOR SEPARATING CELLS USING LOW DENSITY ANTIBODIES AND BALLS |
US4618525A (en) | 1985-06-03 | 1986-10-21 | Minnesota Mining And Manufacturing Company | Coated glass microbubbles and article incorporating them |
US4824776A (en) | 1985-07-25 | 1989-04-25 | Molecular Biosystems, Inc. | Method for increasing the sensitivity of nucleic acid hybridization assays |
US4946778A (en) | 1987-09-21 | 1990-08-07 | Genex Corporation | Single polypeptide chain binding molecules |
US4933447A (en) | 1987-09-24 | 1990-06-12 | Ss Pharmaceutical Co., Ltd. | Quinoline derivatives |
US4957656A (en) | 1988-09-14 | 1990-09-18 | Molecular Biosystems, Inc. | Continuous sonication method for preparing protein encapsulated microbubbles |
US5766849A (en) | 1989-07-11 | 1998-06-16 | Gen-Probe Incorporated | Methods of amplifying nucleic acids using promoter-containing primer sequence |
CA2020958C (en) | 1989-07-11 | 2005-01-11 | Daniel L. Kacian | Nucleic acid sequence amplification methods |
US5427930A (en) | 1990-01-26 | 1995-06-27 | Abbott Laboratories | Amplification of target nucleic acids using gap filling ligase chain reaction |
SE9002480D0 (en) | 1990-07-23 | 1990-07-23 | Hans Lilja | ASSAY OF FREE AND COMPLEXED PROSTATE-SPECIFIC ANTIGEN |
EP0488152A3 (en) | 1990-11-30 | 1992-11-25 | Hitachi, Ltd. | Method for immunoassay and apparatus therefor |
EP0529070A1 (en) | 1991-02-27 | 1993-03-03 | Amoco Corporation | Methods for improving the sensitivity of hybridization assays |
IE920778A1 (en) | 1991-03-12 | 1992-09-23 | Du Pont | Method for specific binding assays using a releasable ligand |
US5665539A (en) | 1991-07-12 | 1997-09-09 | The Regents Of The University Of California | Immuno-polymerase chain reaction system for antigen detection |
US6723303B1 (en) | 1991-09-17 | 2004-04-20 | Amersham Health, As | Ultrasound contrast agents including protein stabilized microspheres of perfluoropropane, perfluorobutane or perfluoropentane |
EP1331011A3 (en) | 1991-10-24 | 2003-12-17 | Isis Pharmaceuticals, Inc. | Derivatized oligonucleotides having improved uptake and other properties |
US5270184A (en) | 1991-11-19 | 1993-12-14 | Becton, Dickinson And Company | Nucleic acid target generation |
US6172208B1 (en) | 1992-07-06 | 2001-01-09 | Genzyme Corporation | Oligonucleotides modified with conjugate groups |
DE4232755A1 (en) | 1992-09-26 | 1994-03-31 | Schering Ag | Microparticle preparations made from biodegradable copolymers |
AU6031094A (en) | 1993-01-15 | 1994-08-15 | Public Health Research Institute Of The City Of New York, Inc., The | Sensitive nucleic acid sandwich hybridization assays and kits |
US5985548A (en) | 1993-02-04 | 1999-11-16 | E. I. Du Pont De Nemours And Company | Amplification of assay reporters by nucleic acid replication |
CA2162568C (en) | 1993-05-10 | 2001-10-30 | Yuichi Oku | Method and reagent for simultaneously assaying one or more ligands in a group of preselected ligands |
US5681697A (en) * | 1993-12-08 | 1997-10-28 | Chiron Corporation | Solution phase nucleic acid sandwich assays having reduced background noise and kits therefor |
US5599677A (en) | 1993-12-29 | 1997-02-04 | Abbott Laboratories | Immunoassays for prostate specific antigen |
US5648213A (en) | 1994-08-30 | 1997-07-15 | Beckman Instruments, Inc. | Compositions and methods for use in detection of analytes |
JPH08178926A (en) | 1994-10-25 | 1996-07-12 | Sumitomo Pharmaceut Co Ltd | Immunoassay plate and use thereof |
WO1996015130A1 (en) | 1994-11-15 | 1996-05-23 | Pharmacia + Upjohn Company | Bicyclic oxazine and thiazine oxazolidinone antibacterials |
SE504798C2 (en) * | 1995-06-16 | 1997-04-28 | Ulf Landegren | Immunoassay and test kits with two reagents that can be cross-linked if adsorbed to the analyte |
DE19524133A1 (en) | 1995-07-03 | 1997-01-09 | Bayer Ag | The use of new and known cationic 4,5-dihydro-1H-1,2,3-triazolium compounds as dyes, new cationic 4,5-dihydro-1H-1,2,3-triazolium compounds and their preparation |
NO301198B1 (en) | 1995-07-14 | 1997-09-22 | Alcatel Kabel Norge As | Cable, process and impregnation pulp |
US5655539A (en) * | 1996-02-26 | 1997-08-12 | Abbott Laboratories | Method for conducting an ultrasound procedure using an ultrasound transmissive pad |
AT403803B (en) | 1996-04-19 | 1998-05-25 | Sanochemia Ltd | NEW BENZAZEPINE DERIVATIVES, THESE MEDICINAL PRODUCTS AND THE USE THEREOF FOR THE PRODUCTION OF MEDICINAL PRODUCTS |
US6165942A (en) | 1996-10-03 | 2000-12-26 | Nissan Chemical Industries, Ltd. | Heterocycle-fused pyrimidinone derivative and herbicidal composition |
US6083484A (en) | 1996-10-17 | 2000-07-04 | Molecular Biosystems, Inc. | Microparticles stabilized by polynuclear chromium complexes and their use as ultrasound contrast agents |
US6245318B1 (en) | 1997-05-27 | 2001-06-12 | Mallinckrodt Inc. | Selectively binding ultrasound contrast agents |
US6086540A (en) | 1997-10-07 | 2000-07-11 | Molecular Biosystems, Inc. | Methods of ultrasound imaging using echogenically persistent contrast agents |
US6531278B1 (en) | 1998-01-14 | 2003-03-11 | Utah State University | Ligand-DNA composition for capture and detection of contaminants on a solid surface |
JP2002512886A (en) | 1998-04-28 | 2002-05-08 | ニユコメド・イメージング・アクシエセルカペト | Improvements in or related to separation methods |
US8063190B2 (en) † | 1998-06-02 | 2011-11-22 | Tom Robin Caine Boyde | Nucleic acid-linked conjugates and methods for making and using them |
US6214566B1 (en) | 1998-11-16 | 2001-04-10 | The Administrators Of The Tulane Educational Fund | Method for detecting anti-squalene antibodies |
JP2003526328A (en) | 1999-01-11 | 2003-09-09 | プレジデント・アンド・フェロウズ・オブ・ハーバード・カレッジ | Isothermal amplification of DNA |
DE60024664T2 (en) * | 1999-10-13 | 2006-07-06 | The University Of British Columbia, Vancouver | METHOD FOR IDENTIFYING COMPOUNDS THAT MODULATE NEURONAL ACTIVITY |
CA2391534A1 (en) * | 1999-11-15 | 2001-05-25 | Drug Innovation & Design, Inc. | Selective cellular targeting: multifunctional delivery vehicles |
US7306904B2 (en) | 2000-02-18 | 2007-12-11 | Olink Ab | Methods and kits for proximity probing |
SE516272C2 (en) † | 2000-02-18 | 2001-12-10 | Ulf Landegren | Methods and kits for analyte detection using proximity probing |
US6511809B2 (en) | 2000-06-13 | 2003-01-28 | E. I. Du Pont De Nemours And Company | Method for the detection of an analyte by means of a nucleic acid reporter |
US7439016B1 (en) | 2000-06-15 | 2008-10-21 | Digene Corporation | Detection of nucleic acids by type-specific hybrid capture method |
JP2004515219A (en) | 2000-06-15 | 2004-05-27 | ボード・オブ・リージェンツ,ザ・ユニヴァーシティ・オヴ・テキサス・システム | Tunable catalytically active nucleic acids |
WO2002068695A2 (en) | 2001-02-22 | 2002-09-06 | Arcturus Engineering, Inc. | Quantitative immunohistochemistry (qihc) |
WO2002083951A1 (en) | 2001-04-10 | 2002-10-24 | Northeastern University | Multiplexed ligand/protein binding assays with pna labels |
EP1249500A1 (en) | 2001-04-12 | 2002-10-16 | chimera biotec GmbH | Method for the determination of an analyte |
WO2003011824A1 (en) | 2001-07-31 | 2003-02-13 | Bristol-Myers Squibb Company | Bicyclic modulators of androgen receptor function |
CN1871517A (en) | 2002-02-19 | 2006-11-29 | 免疫公司 | Methods and reagents for the rapid and efficient isolation of circulating cancer cells |
AU2002310846B2 (en) * | 2002-03-08 | 2008-03-20 | Eidgenossische Technische Hochschule Zurich | Encoded self-assembling chemical libraries (ESACHEL) |
PT1523496E (en) * | 2002-07-18 | 2011-09-29 | Merus B V | Recombinant production of mixtures of antibodies |
KR20050044902A (en) | 2002-08-12 | 2005-05-13 | 다케다 야쿠힌 고교 가부시키가이샤 | Fused benzene derivative and use |
TWI375796B (en) * | 2003-04-18 | 2012-11-01 | Becton Dickinson Co | Immuno-amplification |
US20090176207A1 (en) | 2004-06-23 | 2009-07-09 | Washington University | Methods for Determining Risk of Developing Regular Smoking Behavior |
MX2007003332A (en) | 2004-09-20 | 2007-06-05 | Xenon Pharmaceuticals Inc | Heterocyclic derivatives and their use as stearoyl-coa desaturase inhibitors. |
CN101065497B (en) | 2004-11-03 | 2012-11-21 | 卢卡迪亚技术股份有限公司 | Microbubbles for affinity separation |
KR20070105967A (en) | 2004-11-03 | 2007-10-31 | 아이리스 몰레큘라 다이아그노스틱스, 인코오포레이티드 | Homogeneous analyte detection |
JP4692200B2 (en) | 2005-10-06 | 2011-06-01 | 横河電機株式会社 | Chemical treatment cartridge and method of use thereof |
EP2252893B1 (en) | 2008-02-21 | 2013-10-23 | Iris International, Inc. | Method for early determination of recurrence after therapy for prostate cancer |
-
2005
- 2005-11-03 KR KR1020077012526A patent/KR20070105967A/en not_active Application Discontinuation
- 2005-11-03 CA CA002585781A patent/CA2585781A1/en not_active Abandoned
- 2005-11-03 PT PT05858303T patent/PT1842226E/en unknown
- 2005-11-03 US US11/718,379 patent/US8338579B2/en not_active Expired - Fee Related
- 2005-11-03 AT AT05858303T patent/ATE480643T1/en active
- 2005-11-03 CN CN2005800379854A patent/CN101137758B/en not_active Expired - Fee Related
- 2005-11-03 DE DE602005023529T patent/DE602005023529D1/en active Active
- 2005-11-03 CN CN2012102995602A patent/CN103146808A/en active Pending
- 2005-11-03 JP JP2007539365A patent/JP5188808B2/en not_active Expired - Fee Related
- 2005-11-03 WO PCT/US2005/040133 patent/WO2006137932A2/en active Application Filing
- 2005-11-03 AU AU2005333156A patent/AU2005333156B2/en not_active Ceased
- 2005-11-03 EP EP05858303.0A patent/EP1842226B2/en not_active Not-in-force
- 2005-11-03 DK DK05858303.0T patent/DK1842226T3/en active
- 2005-11-03 ES ES05858303.0T patent/ES2352041T5/en active Active
-
2011
- 2011-05-27 US US13/117,816 patent/US9234890B2/en active Active
-
2012
- 2012-01-25 JP JP2012012830A patent/JP2012120534A/en not_active Withdrawn
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
US4683202B1 (en) | 1985-03-28 | 1990-11-27 | Cetus Corp | |
US5635602A (en) | 1993-08-13 | 1997-06-03 | The Regents Of The University Of California | Design and synthesis of bispecific DNA-antibody conjugates |
US20050026161A1 (en) | 2002-11-01 | 2005-02-03 | Edward Jablonski | Displacement sandwich immuno-PCR |
Non-Patent Citations (2)
Title |
---|
HARLOW; LANE: "Antibodies: A Laboratory Manual", 1988, COLD SPRING HARBOR LABORATORY PRESS |
SAMBROOK ET AL.: "Molecular Cloning: A Laboratory Manual", 1989, COLD SPRING HARBOR LABORATORY PRESS |
Cited By (115)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9234890B2 (en) | 2004-11-03 | 2016-01-12 | Iris International, Inc. | Homogeneous analyte detection |
US8835186B2 (en) | 2004-11-03 | 2014-09-16 | Iris International, Inc. | Microbubbles for affinity separation |
US8513032B2 (en) | 2004-11-03 | 2013-08-20 | Iris International, Inc. | Microbubbles for affinity separation |
CN102016588A (en) * | 2008-02-21 | 2011-04-13 | 艾瑞斯国际有限公司 | Method for early determination of recurrence after therapy for prostate cancer |
JP2011513705A (en) * | 2008-02-21 | 2011-04-28 | アイリス インターナショナル インコーポレイテッド | Method for early determination of recurrence after prostate cancer treatment |
CN103884840A (en) * | 2008-02-21 | 2014-06-25 | 艾瑞斯国际有限公司 | Method for early determination of recurrence after therapy for prostate cancer |
AU2009215799B2 (en) * | 2008-02-21 | 2013-08-22 | Iris International Inc. | Method for early determination of recurrence after therapy for prostate cancer |
WO2009105264A1 (en) * | 2008-02-21 | 2009-08-27 | Iris International Inc. | Method for early determination of recurrence after therapy for prostate cancer |
US9994646B2 (en) | 2009-09-16 | 2018-06-12 | Genentech, Inc. | Coiled coil and/or tether containing protein complexes and uses thereof |
US10619203B2 (en) | 2009-12-15 | 2020-04-14 | Becton, Dickinson And Company | Digital counting of individual molecules by stochastic attachment of diverse labels |
US12060607B2 (en) | 2009-12-15 | 2024-08-13 | Becton, Dickinson And Company | Digital counting of individual molecules by stochastic attachment of diverse labels |
US11993814B2 (en) | 2009-12-15 | 2024-05-28 | Becton, Dickinson And Company | Digital counting of individual molecules by stochastic attachment of diverse labels |
US10202646B2 (en) | 2009-12-15 | 2019-02-12 | Becton, Dickinson And Company | Digital counting of individual molecules by stochastic attachment of diverse labels |
US11970737B2 (en) | 2009-12-15 | 2024-04-30 | Becton, Dickinson And Company | Digital counting of individual molecules by stochastic attachment of diverse labels |
US10106600B2 (en) | 2010-03-26 | 2018-10-23 | Roche Glycart Ag | Bispecific antibodies |
US10381415B2 (en) | 2010-06-16 | 2019-08-13 | Sony Corporation | Display device |
US11081529B2 (en) | 2010-06-16 | 2021-08-03 | Sony Corporation | Display device |
US9761637B2 (en) | 2010-06-16 | 2017-09-12 | Sony Corporation | Display device |
US11871634B2 (en) | 2010-06-16 | 2024-01-09 | Sony Corporation | Display device |
US20160194410A1 (en) * | 2010-12-23 | 2016-07-07 | Roche Diagnostics Operations, Inc. | Binding agent |
WO2012085111A1 (en) | 2010-12-23 | 2012-06-28 | F. Hoffmann-La Roche Ag | Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery |
CN103384681A (en) * | 2010-12-23 | 2013-11-06 | 霍夫曼-拉罗奇有限公司 | Binding agent |
WO2012085113A1 (en) * | 2010-12-23 | 2012-06-28 | Roche Diagnostics Gmbh | Binding agent |
US10982007B2 (en) | 2010-12-23 | 2021-04-20 | Roche Diagnostics Operations, Inc. | Detection of a posttranslationally modified polypeptide by a bivalent binding agent |
US10633460B2 (en) | 2010-12-23 | 2020-04-28 | Roche Diagnostic Operations, Inc. | Binding agent |
US11618790B2 (en) | 2010-12-23 | 2023-04-04 | Hoffmann-La Roche Inc. | Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery |
CN103354840B (en) * | 2011-01-31 | 2016-06-08 | 欧凌科公司 | The contiguous extension that exonuclease can realize is analyzed |
US10731206B2 (en) | 2011-01-31 | 2020-08-04 | Olink Proteomics Ab | Exonuclease enabled proximity extension assays |
JP2014504880A (en) * | 2011-01-31 | 2014-02-27 | オリンク エービー | Proximity extension assay using exonuclease |
EP3323896A3 (en) * | 2011-01-31 | 2018-06-13 | Olink Proteomics AB | Exonuclease enabled proximity extension assays |
US9777315B2 (en) | 2011-01-31 | 2017-10-03 | Olink Proteomics Ab | Exonuclease enabled proximity extension assays |
JP2017108749A (en) * | 2011-01-31 | 2017-06-22 | オリンク プロテオミクス エービー | Proximity extension assay using exonuclease |
WO2012104261A1 (en) | 2011-01-31 | 2012-08-09 | Olink Ab | Exonuclease enabled proximity extension assays |
CN103354840A (en) * | 2011-01-31 | 2013-10-16 | 欧凌科公司 | Exonuclease enabled proximity extension assays |
US9768666B2 (en) | 2011-09-08 | 2017-09-19 | Siemens Aktiengesellschaft | External cooling tube arrangement for a stator of an electric motor |
US9902993B2 (en) | 2012-01-30 | 2018-02-27 | Olink Proteomics Ab | Hyperthermophilic polymerase enabled proximity extension assay |
WO2013113699A3 (en) * | 2012-01-30 | 2014-03-20 | Olink Ab | Hyperthermophilic polymerase enabled proximity extension assay |
US9688758B2 (en) | 2012-02-10 | 2017-06-27 | Genentech, Inc. | Single-chain antibodies and other heteromultimers |
US11579327B2 (en) | 2012-02-14 | 2023-02-14 | American Science And Engineering, Inc. | Handheld backscatter imaging systems with primary and secondary detector arrays |
US11634708B2 (en) | 2012-02-27 | 2023-04-25 | Becton, Dickinson And Company | Compositions and kits for molecular counting |
US10941396B2 (en) | 2012-02-27 | 2021-03-09 | Becton, Dickinson And Company | Compositions and kits for molecular counting |
US10106612B2 (en) | 2012-06-27 | 2018-10-23 | Hoffmann-La Roche Inc. | Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof |
WO2014001326A1 (en) | 2012-06-27 | 2014-01-03 | F. Hoffmann-La Roche Ag | Method for the selection and production of tailor-made, selective and multi-specific therapeutic molecules comprising at least two different targeting entities and uses thereof |
US11407836B2 (en) | 2012-06-27 | 2022-08-09 | Hoffmann-La Roche Inc. | Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof |
US11421022B2 (en) | 2012-06-27 | 2022-08-23 | Hoffmann-La Roche Inc. | Method for making antibody Fc-region conjugates comprising at least one binding entity that specifically binds to a target and uses thereof |
US9862779B2 (en) | 2012-09-14 | 2018-01-09 | Hoffmann-La Roche Inc. | Method for the production and selection of molecules comprising at least two different entities and uses thereof |
US10927419B2 (en) | 2013-08-28 | 2021-02-23 | Becton, Dickinson And Company | Massively parallel single cell analysis |
US11702706B2 (en) | 2013-08-28 | 2023-07-18 | Becton, Dickinson And Company | Massively parallel single cell analysis |
US10954570B2 (en) | 2013-08-28 | 2021-03-23 | Becton, Dickinson And Company | Massively parallel single cell analysis |
US10131958B1 (en) | 2013-08-28 | 2018-11-20 | Cellular Research, Inc. | Massively parallel single cell analysis |
US11618929B2 (en) | 2013-08-28 | 2023-04-04 | Becton, Dickinson And Company | Massively parallel single cell analysis |
US10151003B2 (en) | 2013-08-28 | 2018-12-11 | Cellular Research, Inc. | Massively Parallel single cell analysis |
US11926865B2 (en) | 2014-06-23 | 2024-03-12 | The Board Of Trustees Of The Leland Stanford Junior University | On-slide staining by primer extension |
US11299770B2 (en) | 2014-06-23 | 2022-04-12 | The Board Of Trustees Of The Leland Stanford Junior University | On-slide staining by primer extension |
US11634753B2 (en) | 2014-06-23 | 2023-04-25 | The Board Of Trustees Of The Leland Stanford Junior University | On-slide staining by primer extension |
US10982263B2 (en) | 2014-06-23 | 2021-04-20 | The Board Of Trustees Of The Leland Stanford Junior University | On-slide staining by primer extension |
NL2013857B1 (en) * | 2014-11-21 | 2016-10-11 | Piculet Biosciences Tech B V | Self-assembled bivalent ligand complex (SABLC) libraries and methods for screening such libraries. |
WO2016080838A1 (en) * | 2014-11-21 | 2016-05-26 | Piculet Biosciences Technologies B.V. | Self-assembled bivalent ligand complex (sablc) libraries and methods for screening such libraries |
US11999801B2 (en) | 2014-12-03 | 2024-06-04 | Hoffman-La Roche Inc. | Multispecific antibodies |
US10633457B2 (en) | 2014-12-03 | 2020-04-28 | Hoffmann-La Roche Inc. | Multispecific antibodies |
USRE48913E1 (en) | 2015-02-27 | 2022-02-01 | Becton, Dickinson And Company | Spatially addressable molecular barcoding |
US11300703B2 (en) | 2015-03-20 | 2022-04-12 | Rapiscan Systems, Inc. | Hand-held portable backscatter inspection system |
US11561320B2 (en) | 2015-03-20 | 2023-01-24 | Rapiscan Systems, Inc. | Hand-held portable backscatter inspection system |
US11535882B2 (en) | 2015-03-30 | 2022-12-27 | Becton, Dickinson And Company | Methods and compositions for combinatorial barcoding |
US11390914B2 (en) | 2015-04-23 | 2022-07-19 | Becton, Dickinson And Company | Methods and compositions for whole transcriptome amplification |
US11332776B2 (en) | 2015-09-11 | 2022-05-17 | Becton, Dickinson And Company | Methods and compositions for library normalization |
US10781473B2 (en) | 2015-10-21 | 2020-09-22 | Olink Proteomics Ab | Method for generating proximity probes |
US11220685B2 (en) | 2016-05-31 | 2022-01-11 | Becton, Dickinson And Company | Molecular indexing of internal sequences |
US11525157B2 (en) | 2016-05-31 | 2022-12-13 | Becton, Dickinson And Company | Error correction in amplification of samples |
US10640763B2 (en) | 2016-05-31 | 2020-05-05 | Cellular Research, Inc. | Molecular indexing of internal sequences |
CN106124781A (en) * | 2016-07-07 | 2016-11-16 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of method based on aptamers sandwich fluorescent quenching technology for detection demethyl testosterone |
US11168350B2 (en) | 2016-07-27 | 2021-11-09 | The Board Of Trustees Of The Leland Stanford Junior University | Highly-multiplexed fluorescent imaging |
US10338066B2 (en) | 2016-09-26 | 2019-07-02 | Cellular Research, Inc. | Measurement of protein expression using reagents with barcoded oligonucleotide sequences |
US11467157B2 (en) | 2016-09-26 | 2022-10-11 | Becton, Dickinson And Company | Measurement of protein expression using reagents with barcoded oligonucleotide sequences |
US11782059B2 (en) | 2016-09-26 | 2023-10-10 | Becton, Dickinson And Company | Measurement of protein expression using reagents with barcoded oligonucleotide sequences |
US11460468B2 (en) | 2016-09-26 | 2022-10-04 | Becton, Dickinson And Company | Measurement of protein expression using reagents with barcoded oligonucleotide sequences |
US11319583B2 (en) | 2017-02-01 | 2022-05-03 | Becton, Dickinson And Company | Selective amplification using blocking oligonucleotides |
EP3580356B1 (en) * | 2017-02-09 | 2021-03-31 | Atlas Antibodies AB | Method for determining levels of interactions between biomolecules |
US10669570B2 (en) | 2017-06-05 | 2020-06-02 | Becton, Dickinson And Company | Sample indexing for single cells |
US12084712B2 (en) | 2017-06-05 | 2024-09-10 | Becton, Dickinson And Company | Sample indexing for single cells |
WO2018226293A1 (en) * | 2017-06-05 | 2018-12-13 | Becton, Dickinson And Company | Sample indexing for single cells |
US10676779B2 (en) | 2017-06-05 | 2020-06-09 | Becton, Dickinson And Company | Sample indexing for single cells |
US11773441B2 (en) | 2018-05-03 | 2023-10-03 | Becton, Dickinson And Company | High throughput multiomics sample analysis |
US11365409B2 (en) | 2018-05-03 | 2022-06-21 | Becton, Dickinson And Company | Molecular barcoding on opposite transcript ends |
US11525930B2 (en) | 2018-06-20 | 2022-12-13 | American Science And Engineering, Inc. | Wavelength-shifting sheet-coupled scintillation detectors |
US11639517B2 (en) | 2018-10-01 | 2023-05-02 | Becton, Dickinson And Company | Determining 5′ transcript sequences |
US11932849B2 (en) | 2018-11-08 | 2024-03-19 | Becton, Dickinson And Company | Whole transcriptome analysis of single cells using random priming |
US11492660B2 (en) | 2018-12-13 | 2022-11-08 | Becton, Dickinson And Company | Selective extension in single cell whole transcriptome analysis |
US11661631B2 (en) | 2019-01-23 | 2023-05-30 | Becton, Dickinson And Company | Oligonucleotides associated with antibodies |
US12071617B2 (en) | 2019-02-14 | 2024-08-27 | Becton, Dickinson And Company | Hybrid targeted and whole transcriptome amplification |
US11965208B2 (en) | 2019-04-19 | 2024-04-23 | Becton, Dickinson And Company | Methods of associating phenotypical data and single cell sequencing data |
US11939622B2 (en) | 2019-07-22 | 2024-03-26 | Becton, Dickinson And Company | Single cell chromatin immunoprecipitation sequencing assay |
US11773436B2 (en) | 2019-11-08 | 2023-10-03 | Becton, Dickinson And Company | Using random priming to obtain full-length V(D)J information for immune repertoire sequencing |
US11649497B2 (en) | 2020-01-13 | 2023-05-16 | Becton, Dickinson And Company | Methods and compositions for quantitation of proteins and RNA |
WO2021191449A1 (en) | 2020-03-27 | 2021-09-30 | Olink Proteomics Ab | Method for detecting analytes |
GB202004469D0 (en) | 2020-03-27 | 2020-05-13 | Olink Proteomics Ab | Controls for proximity detection assays |
EP4269608A1 (en) | 2020-03-27 | 2023-11-01 | Olink Proteomics AB | Proximity detection assays |
GB202004472D0 (en) | 2020-03-27 | 2020-05-13 | Olink Proteomics Ab | Method for detecting analytes of varying abundance |
DE202021004362U1 (en) | 2020-03-27 | 2024-01-11 | Olink Proteomics Ab | Controls for proximity detection assays |
WO2021191448A1 (en) | 2020-03-27 | 2021-09-30 | Olink Proteomics Ab | Method for detecting analytes |
WO2021191450A1 (en) | 2020-03-27 | 2021-09-30 | Olink Proteomics Ab | Controls for proximity detection assays |
WO2021191442A1 (en) | 2020-03-27 | 2021-09-30 | Olink Proteomics Ab | Method for detecting analytes of varying abundance |
US11661625B2 (en) | 2020-05-14 | 2023-05-30 | Becton, Dickinson And Company | Primers for immune repertoire profiling |
US11932901B2 (en) | 2020-07-13 | 2024-03-19 | Becton, Dickinson And Company | Target enrichment using nucleic acid probes for scRNAseq |
US20220119552A1 (en) * | 2020-10-15 | 2022-04-21 | City Of Hope | Complementary rna linked bispecific t-cell engaging antibodies |
US12043673B2 (en) * | 2020-10-15 | 2024-07-23 | City Of Hope | Complementary RNA linked bispecific T-cell engaging antibodies |
WO2022101414A1 (en) | 2020-11-12 | 2022-05-19 | Oslo Universitetssykehus Hf | Method of determining dlbcl prognosis |
US11739443B2 (en) | 2020-11-20 | 2023-08-29 | Becton, Dickinson And Company | Profiling of highly expressed and lowly expressed proteins |
US11340361B1 (en) | 2020-11-23 | 2022-05-24 | American Science And Engineering, Inc. | Wireless transmission detector panel for an X-ray scanner |
US11726218B2 (en) | 2020-11-23 | 2023-08-15 | American Science arid Engineering, Inc. | Methods and systems for synchronizing backscatter signals and wireless transmission signals in x-ray scanning |
GB202018503D0 (en) | 2020-11-25 | 2021-01-06 | Olink Proteomics Ab | Analyte detection method employing concatamers |
WO2022112300A1 (en) | 2020-11-25 | 2022-06-02 | Olink Proteomics Ab | Analyte detection method employing concatemers |
WO2024100258A1 (en) | 2022-11-11 | 2024-05-16 | Olink Proteomics Ab | Library of proximity probes and method of use |
EP4382534A1 (en) | 2022-12-06 | 2024-06-12 | Biomérieux | Synthetic peptide for use in an immunomolecular assay |
WO2024121128A1 (en) | 2022-12-06 | 2024-06-13 | bioMérieux | Synthetic peptide for use in an immunomolecular assay |
Also Published As
Publication number | Publication date |
---|---|
WO2006137932A3 (en) | 2007-09-20 |
US8338579B2 (en) | 2012-12-25 |
AU2005333156A1 (en) | 2006-12-28 |
JP2012120534A (en) | 2012-06-28 |
JP5188808B2 (en) | 2013-04-24 |
CA2585781A1 (en) | 2006-12-28 |
ES2352041T5 (en) | 2014-12-22 |
CN103146808A (en) | 2013-06-12 |
EP1842226B2 (en) | 2014-07-02 |
DK1842226T3 (en) | 2010-10-18 |
US20120082988A1 (en) | 2012-04-05 |
ES2352041T3 (en) | 2011-02-15 |
WO2006137932A9 (en) | 2007-02-15 |
EP1842226A2 (en) | 2007-10-10 |
DE602005023529D1 (en) | 2010-10-21 |
EP1842226A4 (en) | 2009-08-05 |
CN101137758B (en) | 2012-10-10 |
US9234890B2 (en) | 2016-01-12 |
AU2005333156B2 (en) | 2011-05-26 |
EP1842226B1 (en) | 2010-09-08 |
KR20070105967A (en) | 2007-10-31 |
CN101137758A (en) | 2008-03-05 |
JP2008518605A (en) | 2008-06-05 |
US20080131883A1 (en) | 2008-06-05 |
PT1842226E (en) | 2010-11-29 |
ATE480643T1 (en) | 2010-09-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US8338579B2 (en) | Homogeneous analyte detection | |
AU783644B2 (en) | Methods and kits for proximity probing | |
US7306904B2 (en) | Methods and kits for proximity probing | |
EP1563100B1 (en) | Displacement sandwich immuno-pcr | |
EP1815028B1 (en) | Microbubbles for affinity separation | |
EP0339686A1 (en) | Nucleic acid hybridization assay | |
US20150044666A1 (en) | Non-equilibrium two-site assays for linear, ultrasensitive analyte detection | |
US8927210B2 (en) | Conjugate complexes for analyte detection | |
AU2011213911A1 (en) | Homogeneous analyte detection | |
KR20220160093A (en) | Methods for detecting analytes of varying abundance | |
JPH06235727A (en) | Biological information converting element and biological species detecting method employing the same |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
WWE | Wipo information: entry into national phase |
Ref document number: 200580037985.4 Country of ref document: CN |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2007539365 Country of ref document: JP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2005333156 Country of ref document: AU |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2585781 Country of ref document: CA |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 3538/DELNP/2007 Country of ref document: IN |
|
ENP | Entry into the national phase |
Ref document number: 2005333156 Country of ref document: AU Date of ref document: 20051103 Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2005858303 Country of ref document: EP Ref document number: 1020077012526 Country of ref document: KR |
|
WWP | Wipo information: published in national office |
Ref document number: 2005858303 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 11718379 Country of ref document: US |