肌钙蛋白手册

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1 心肌肌钙蛋白 I

2 HYTEST 心肌肌钙蛋白I 目录介绍 3 特异性识别不同位点的心肌肌钙蛋白I抗体 4 超敏心肌肌钙蛋白概念 5 心肌肌钙蛋白I检测的影响因素 6 心肌肌钙蛋白I免疫检测系统的建立与抗体配对 10 外周循环中心肌肌钙蛋白I的异质性与检测标准化 13 蛋白免疫印迹检测心肌肌钙蛋白I及其片段 14 识别不同物种心肌肌钙蛋白I的抗体 14 心肌肌钙蛋白I单体与复合物 16 心肌肌钙蛋白T 18 肌钙蛋白C 20 参考文献 21 HyTest科学家的精品肌钙蛋白文章 22 订购信息 24 Abbreviations AMI cc ctni ctnt HAMA hs-ctn MAb sktni sktnt Tn TnC 2 Acute myocardial infarction cell culture; produced in vitro (in the Cat or MAb name) cardiac troponin I cardiac troponin T human anti-mouse antibody high-sensitivity cardiac troponin monoclonal antibody skeletal troponin I skeletal troponin T Troponin Troponin C

3 T TnT I TnI C TnC I C TnI TnT 2080cTnI ctnt sktni 骼 sktnt I 骼 T I 骼 TI ctni AMI6-9 T ctntctni 2 ctnt 骼 3 3

4 HYTEST I I HyTestcTnI 20cTnI ctni ctni 1 HAMA 801 M M F4 P4-9F C7 P4-14G5 4C2 19C ADGSSDAAREPRPAPAPIRRRSSNYRAYATEPHAKKKSKISASRKLQLKTLLLQIAKQELEREAEERRGEKGRALSTRCQ A E10 16A11 17F M PLELAGLGFAELQDLCRQLHARVDKVDEERYDIEAKVTKNITEIADLTQKIFDLRGKFKRPTLRRVRISADAMMQALLGA MF4 458 C5 625 p RAKESLDLRAHLKQVKKEDTEKENREVGDWRKNIDALSGMEGRKKKFES 1. HyTest ctni 30cTnI HyTestcTnI ELISA Western Blotting 4

5 ctni ctnt 1987 ng/ml(μ g/l) (10 ng/l vs.10 ng/ml) 3-6 AMI pg/ml ng/l 1-3 AMI AMI % 2. ctn50% 5

6 HYTEST I I ctni ctni ctni ctni ctni ctni TnI ctni 2 ctni N stni ctni stni stni ctni 52% 46% ctni stni ctni NH 2 COOH I IcTnI ctni stni0.1% 6

7 ctni ctni ctni ctni ctni ctni ctni ctni ctnitnc Sites of phosphorylation TnC TnT Sites of proteolysis Heparin Tnl Autoantibodies 3. ctni ctni TnC ctnt ctni I-T-C TnC ctni ctni I-CI-T-C TnC ctni TnC ctni ctni 7

8 HYTEST I ctni ctni TnC % of ctni immunological activity ctni TnC ctni NC C7- M18 19C C7-19C7- M C7+MF4 M18- MF4 0 h 170 h 4. 8T ctni M18-MF4 ctni ctni 22B AMI ctni 4 ctni 18 ctni ctni % of ctni immunological activity Heparin +Heparin 80 AMI ctni C7- M C7 19C C7+MF4 19C7 5. 5IU/ ng/m- L 228 8

9 9 0 CPS Buffer Sample 1 Sample 2 Sample 3 ctni HAMA HAMA HAMA HAMA AMI 25 ctnihama cut-off AMI ctni MI HAMA 5.5% HAMA14% 26 HAMA RecChim16A11 HAMA HAMA 6 6. HAMA 19C7-16A11 3 HAMAHAMA 807ng/mL 1388ng/mL 6220ng/mL BUFFER

10 HYTEST I ctni ctni ctni 1 AMI ctni 1. ctni Assay type Capture Detection 19C7cc 16A11cc 19C7cc 560cc C7cc 560cc 458 4C2cc 19C7cc C7cc + MF4cc 7B9cc (specific to TnC) cc 19C7cc + MF4cc cc 19C7cc + MF4cc cc 19C7cc + MF4cc M cc 19C7cc + MF4cc 10

11 ctni ctni Assay type Capture Detection 20C6 (specific to ctn complex) 560cc 560cc 20C C6 7B9cc (specific to TnC) 19C7cc 560cc Assay type MAb1 MAb2 MAb3 4C2cc 7B9cc 20C6 19C7cc 560cc 20C or cc 7B9cc 20C6 19C7cc 267 4T21/2 19C7cc 560cc 4T21/2 Assay type MAb1 MAb2 MAb3 MAb C2cc 560cc 7B9cc 20C6 11

12 12 ctni TnC ctni TnC ctni ctnt HYTEST I HyTest ctni ctni-tnc 17,27,28cTnI 28cTn ctnitnc 1 TnC 1 ctni7 TnC TnC 4T27 4T27cc ctnictn 1 ctn 1 ctni TnC ctn 4TC2 ctni 7. TnC ctni TnI TnC 8. TnC ctni ctn ctni

13 ctni ctni ctni ctnisrm 2921 NIST HyTest 1000 ctni RESEARCH C OMMERCIAL A SS A Y S A SSA Y S Abbott AxSYM ADV Abbott Architect Abbott Architect STAT hs-ctni Abbott i-stat Alere Triage SOB* Alere Triage Cardio 3 Beckman Coulter Access Accu biomerieux Vidas Ultra Mitsubishi PATHFAST ctni Mitsubishi PATHFAST ctni-ii Ortho VITROS Troponin I ES Radiometer AQT90 FLEX TnI Response Biomedical RAMP Roche E 2010/cobas e 411/Roche E 170/cobas e 601/602 ctni Siemens ADVIA Centaur TnI-Ultra Siemens Dimension EXL TNI Siemens Dimension RxL CTNI Siemens Dimension VISTA CTNI Siemens IMMULITE 1000 Turbo Siemens IMMULITE 1000 Siemens IMMULITE 2000 XPi Siemens Stratus CS ctni Tosoh ST AIA-PACK Beckman Coulter Access hs-ctni Singulex Erenna hs-ctni * Data not available for the capture antibody 9. ctni IFCC (Troponin assay analytical characteristics, version Octobe 2013) 13

14 HYTEST I ctni ctni WB ctni WB 19C7 16A11 MF4 ctni ctni ctni M155-19C7 ctniwb ctni 3 14

15 3. WB ctnictni MAb Human Bovine Porcine Goat Canine Rabbit Cat Rat Mouse Fish 4C C E A C MF B N/A 10F N/A 4. ctni Coating MAb Detecting MAb 19C7 M155 MF4 B C M R Rb B C M R Rb B C M R 9F6 4C2 801 M155 19C7 625 MF4 B C MRRb CPS 100 Human ctni (in native complex) Rat ctni (in native complex) Canine ctni (in native complex) 10. Mouse ctni (in native complex) M15519C7 10 0,01 0, ctni concentration (ng/ml) 15

16 kda I kda HYTEST I HyTestcTnI20 I ctni 8T53 ctni 5%cTnI ctni N ctni SDS-PAGE ctni 29KDa11 ctni 8T53ph 8T53dp 11. ctni 1μg ctni 8T % SDS-PAGE 12. ctni 1μg c TnI 8RTI710-20% SDS-PAGE I ctni 8RTI7 TNNI3 N SDS-PAGE 29KDa11 ctni

17 I-C I-T-C ctni TnC 100 Native ctn complex 80 EDTA 60 % of ctni immunological activity ctni I-C I-T-C 40 Native ctni ctnii-c I-T-C I-T-CcTnI ctni 4 Incubation time (days) ctni 8T62c AMI L4 ctni ctni ctni SDS-PAGE 14 ctnt ctni TnC KDa AMI ctni ctni 14. 1μg 15 8T % SDS-PAGE kda HyTestI-T-C AACC ctni NISTwww.nist.gov 17

18 HYTEST I T ctnt ctnt TNNT2 ctnt ctntmrna ctnt 4T19cc LoD 0.3ng/L ctni TnT 6 TnT3 30μg/mLcTnT ctnt KDa ctnt ctni ctnt 15 7G7* 9G6* 7F4* MSDIEEVVEEYEEEEQEEAAVEEQEEAAEEDAEAEAETEETRAEEDEEEEEAKEAEDGPMEESKPKPRSFMPNLVPPKIPDGERVDFDDIHRKRMEKDLN 1F11* 329cc 1A11* 300cc 2F3* 406cc 1C11cc* ELQALIEAHFENRKKEEEELVSLKDRIERRRAERAEQQRIRNEREKERQNRLAEERARREEEENRRKAEDEARKKKALSNMMHFGGYIQKQAQTERKSGK 7E7* RQTEREKKKKILAERRKVLAIDHLNEDQLREKAKELWQSIYNLEAEKFDLQEKFKQQKYEINVLRNRINDNQKVSKTRGKAKVTGRWK 15. ctnt ctnt 18

19 80 AMI 329cc-406cc 406cc-300cc 16 ctnt HyTest ctnt assay, ng/l Commercial hs-ctnt assay, ng/l HyTest ctnt assay, ng/l Commercial hs-ctnt assay, ng/l 16. HyTest 329cc-406cc 406cc-300cc HyTest ctnt ctnt 5 5. WBcTnTcTnT MAb Human Bovine Porcine Goat Canine Rabbit Cat Rat Mouse Fish 7F4 ++ N/A ++ N/A N/A N/A - 1F G F A G F A

20 20 C TnC kda kda TnC HYTEST I 17. ctni 1μg ctnt 8T % SDS-PAGE 18. ctnt 1μg c TnT 8RTT510-20% SDS-PAGE ctnt HyTestcTnT 8T13 SDS-PAGE ctnt 39KDa17 ctnt ctnt 6TnT3 ctnt ctnt 8RTT5 288 TnT3N SDS-PAGE ctnt 39KDa 18 TnT TnT 骼 TnT 8RST2 骼 TnT 8RFT4 ctnt TnC TnC KDa4.05 TnC TnI AMIcTnITnC ctni ctni HyTestTnC 8T57 SDS-PAGE ctnc 18KDa TnC ctni

21 1) Gomes, AV et al. The role of Troponin in muscle contraction. Life. 2002, 54: ) Bodor, GS et al. Cardiac troponin-i is not expressed in fetal and healthy or diseased adult human skeletal muscle tissue; Clin Chem. 1995, 41(12): ) Jaffe, AS et al. Diseased skeletal muscle. A noncardiac source of increased circulating concentrations of cardiac troponin T; J Am Coll Cardiol. 2011, 58(17): ) Cummins, B et al. Cardiac-specifc troponin-i radioimmunoassay in the diagnosis of acute myocardial infarction. Am Heart J. 1987, 113(6): ) Katus, HA et al. Enzyme linked immunoassay of cardiac troponin T for the detection of acute myocardial infarction in patients. J Mol Cell Cardiol. 1989, 21 (12): ) Myocardial infarction redefned A consensus document of The Joint European Society of Cardiology/ American College of Cardiology Committee for the Redefnition of Myocardial Infarction Eur. Heart J. 2000, 21(18): ) Thygesen, K et al. Third universal defnition of myocardial infarction. Circulation. 2012, 126: ) Sandoval, Y and Thygesen, K. Myocardial Infarction Type 2 and Myocardial Injury. Clin Chem. 2017, 63 (1): ) Cullen, L et al. Early rule-out and rule-in strategies for myocardial infarction. Clin Chem. 2017, 63(1): ) Diercks, DB et al. Diagnostic accuracy of a point-of-care troponin I assay for acute myocardial infarction within 3 hours after presentation in early presenters to the emergency department with chest pain. Am Heart J. 2012, 163: ) Reichlin, T et al. One-Hour Rule-out and Rule-in of Acute Myocardial Infarction Using High-Sensitivity Cardiac Troponin T. Arch Intern Med. 2012, 172(16): ) Apple, FS et al., on behalf of the IFCC Task Force4 on Clinical Applications of Cardiac Bio-Markers. IFCC educational materials on selected analytical and clinical applications of high-sensitivity cardiac troponin assays. Clin Biochem. 2015, 48: ) Sandoval, Y et al. Diagnosis of type 1 and type 2 myocardial infarction using a high-sensitivity cardiac troponin I assay with gender-specifc 99th percentiles based on the Third Universal Defnition of Myocardial Infarction classifcation system. Clin Chem. 2015, 61: ) Sandoval, Y et al. Present and future of high sensitivity cardiac troponin in clinical practice: a paradigm shift to high sensitivity assays. Amer J Med. 2016, 129: ) Apple, FS et al., for the IFCC Task Force on Clinical Applications of Cardiac Bio-Markers. Cardiac troponin assays: guide to understanding analytical characteristics and their impact on clinical care, Clin Chem. 2017, 63(1): ) Prof. Fred S. Apple, personal communication. 17) Katrukha, AG et al. Degradation of cardiac troponin I: implication for reliable immunodetection. Clin Chem. 1998, 44(12): ) Zhang, J et al. Top-down quantitative proteomics identifed phosphorylation of cardiac troponin I as a candidate biomarker for chronic heart failure. J Proteome Res. 2011, 10(9): ) Katrukha, AG et al. Biochemical factors influencing measurement of cardiac troponin I in serum. Clin Chem Lab Med. 1999, 37(11-12): ) Eriksson, S et al. Negative interference in cardiac troponin I immunoassays from a frequently occurring serum and plasma component. Clin Chem. 2003, 49 (7): ) Eriksson, S et al. Autoantibodies against cardiac troponins. N Engl J Med. 2005, 352(1): ) Eriksson, S et al. Negative interference in cardiac troponin I immunoassays by circulating troponin autoantibodies. Clin Chem. 2005, 51(5): ) Rahman, A and Broadley, SA. Review article: Elevated troponin: diagnostic gold or fool s gold? Emerg Med Australas. 2014, 26: ) Morton, A. When lab tests lie heterophile antibodies. Aust Fam Physician 2014, 43, ) Zaidi, A and Cowell, R. False positive cardiac troponin elevation due to heterophile antibodies - more common than we recognise. BMJ Case Reports, 2010, Jul. 26) Fleming, SM et al. False-positive cardiac troponin I in a routine clinical population. Am J Cardiol. 2002, 89, ) Wu, AHB. Analytical issues for clinical use of cardiac troponin. Cardiovascular biomarkers. Editor: Morrow DA. Totowa (NJ): Humana Press; P ) Bates, KJ et al. Circulating immunoreactive cardiac troponin forms determined by gel filtration chromatography after acute myocardial infarction. Clin Chem. 2010, 56:

22 HYTEST I HyTest Katrukha AG, Bereznikova AV, Esakova TV, 31 Filatov VL, Bulargina TV and Gusev NB. A new method of human cardiac troponin I ctni ctni and troponin T purifcation. Biochem. Mol. Biol. Int. 1995, 36: Katrukha AG, Bereznikova AV, Filatov VL, Esakova TV, Kolosova OV, Pettersson K, Lövgren T, Bulargina I T TV, Trifonov IR, Gratsiansky NA, Pulkki K, Voipio-Pulkki LM and C5 Gusev I NB. Degradation of cardiac troponin I: implication for reliable immunodetection. Katrukha AG, Bereznikova AV, Esakova TV, Pettersson K, Lövgren T, Severina ME, Pulkki K, Vuopio-Pulkki LM and Gusev NB. Troponin I is released in bloodstream of patients with acute myocardial infarction not in free form but as complex. Clin. Chem. 1997, 43 (8): Clin. Chem. 1998, 44(12): C C- 3 ctni ctni Filatov VL, Katrukha AG, Bulargina TV and TnC Gusev ctni NB. Troponin: structure, properties, ctni and mechanism of functioning.biochemistry. 1999, 64(9): ctni TnC ctni Filatov VL, Katrukha AG, Bereznikova AV, T Esakova TV, Bulargina TV, Kolosova OV, Katrukha A, Bereznikova A, Filatov V and Severin ES and Gusev NB.Epitope mapping Esakova T. Biochemical factors influencing of anti-troponin I monoclonal antibodies. measurement of cardiac troponin I in Biochem. Mol. Biol. Int. 1998, 45(6): serum. Clin. Chem. Lab. Med. 1999, 37(11- ctni 12): SPOT 22

23 Vylegzhanina AV, Kogan AE, Katrukha IA, Antipova OV, Kara AN, Bereznikova AV, Koshkina EV, Katrukha AG. Anti Cardiac Troponin Autoantibodies Are Specifc to the Conformational Epitopes Formed by Cardi- ac Troponin ctni I and Troponin T in the Ternary Troponin Complex. Clin. ctni Chem. 2017, 63(1), ctni Katrukha A, Bereznikova A and Pettersson K. New approach to standardization of human TnI-T-C cardiac troponin I (ctni). Scand. J. Clin. Lab. ctni AMI Invest., Suppl. 1999, 230: TnI-T-C ctni 6 ctni 21 ctni Katrukha IA, Kogan AE, Vylegzhanina AV, Serebryakova MV, Koshkina EV, Bereznikova AV, Katrukha AG. Thrombin-Mediated Degradation of Human Cardiac Troponin T. Clin Chem. 2017, 63(6): Katrukha AG. Antibody selection strategies in cardiac troponin assays. Cardiac Markers, 2003, 2nd edition, Edited by Alan HB. Wu ctnt 29KDa I T Vylegzhanina AV, Katrukha IA, Kogan AE and Bereznikova AV. Epitope Specifcity of Anti Cardiac Troponin I Monoclonal Antibody 8I-7. Clin. Chem. 2013, 59(12): ctni I mab 8I-7 I HyTestI US EP

24 HYTEST I I TnI I 4T21 P4-14G5 P4-9F M C2 3C7 228 M F4 19C A12 8E10 16A11 17F M46 IgG3 IgG3 IgG2a IgG2a IgG2b 4T21cc ctni RC4T21 I 4T45 I 4T46 4TC2 I 4T C5 MF4 p C2cc M155cc 19C7cc 560cc 16A12cc 16A11cc MF4cc RecChim19C7 RecChim16A11 1G11 22B11 20C6 Tcom8 12F10 7G2 IgM IgG2a IgG2b IgG2a IgG2b IgG2b IgG2b IgG2b IgG2b IgG2b EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r a.a.r , ctni EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r , TnI <10 % EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r , TnI <10 % EIA, WB, a.a.r , TnI <10 % EIA, WB, a.a.r , TnI >50 % EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA EIA EIA, WB, a.a.r. N/A EIA, WB, a.a.r EIA EIA EIA, WB EIA, WB 24

25 I TnI I 4T21/2 EIA I 8T53 I8RTI7 I8T53dp I (I-C) (I-T-C), 8T53ph 8IC63 8T62 (I-T-C), 8T62a I 8T60 I K01 骼 I, 8T25 N/A N/A N/A N/A N/A ng/ml I 骼 I I I I I I (I-T-C), (I-T-C), (I-T-C), (I-T-C), (I-T-C), (I-T-C), I, I, I, I, I, I, 8T53b 8T53c 8T53m 8T53p 8T53rb 8T53r 8T62b 8T62c 8T62m 8T62p 8T62rb 8T62r 8T25b 8T25c 8T25m 8T25p 8T25rb 8T25r N/A N/A N/A N/A N/A N/A 骼骼骼骼骼骼 8TFS I 9CT4 4C2cTnI 25

26 HYTEST I T TnT T T, 4T19 4T19cc 9G6 7F4 7G7 2F3 1A11 1F11 1C11 7E7 1C11cc 300cc 329cc 406cc 9G6 7F4 7G7 2F3 1A11 1F11 1C11 7E7 1C11cc 300cc 329cc 406cc EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, WB, a.a.r EIA, a.a.r EIA, a.a.r EIA, a.a.r T 8T13 T8RTT5 T T T (I-T-C), 8T24 8RFT4 8RST2 8T62 N/A (I-T-C), 8T62a N/A T T T T T T T, T, T, T, T, T, 8T13b 8T13c 8T13m 8T13p 8T13rb 8T13r 8T24b 8T24c 8T24m 8T24p 8T24rb 8T24r T 4T19/2 EIA 26

27 C C, 4T27 4T27cc 1A2 7B9 7B9cc IgG2b EIA, WB EIA, WB EIA, WB 4TC2 20C6 Tcom8 IgG2b EIA EIA C 8T57 / C 8RSC4 2C 8RKC3 >90% (I-C), (I-T-C), 8IC63 8T62 N/A N/A (I-T-C), 8T62a N/A C C C C C C 8T57b 8T57c 8T57m 8T57p 8T57rb 8T57r 27

28 Together. Today and Tomorrow. i hytestchina@hytest.fi

Cardiac Troponin I

Cardiac Troponin I website:wwwsupermtcomtw 1 ? (AMI) (cardiac angina) [ ][WHO1971] ECG ST Q [ CK MB CK-MB LD-1] [Ellis,1991] ( ) (CK-MB ) CK-MB (ECG) % (silent infarction) (silent ischemia) 20%30% 4% Cardiac STAus TM (CK-MBMyoglobulinTroponin