The Preparation Of The Anti-cTnI Antibody And Disscussion Of Detection Method For CTnI

4h, cTnI then appears after the acute cardiac arrest in the blood, continues at least 4 day, forms the long time window.Because cTnI these nature and the unique amino acid sequence cause it to become time the cardiac muscle cell damage designated object which pays attention.In the acute heart stem diagnosis aspect, cTnI surpasses CK, CKMB, the myo- red-painted egg waits in vain the traditional cardiac muscle cell damage designated object, the cardiac muscle cell damage designated object which its biggest merit lies in its cardiac muscle specificity, cTnI is the only one kind in which the damage regenerates the skeletal muscle expresses not, even if in the concomitance skeletal muscle damage situation (flesh wound, bruise, heart surgery and so on) also can accurately discover the heart stem promptly.Troponin I,the inhibitory protein of troponin-tropomyosin complex, exists three isotype forms I fast and slow skeletal and cardiac muscles that are encoded by three separate genes. The amino acid sequences of the two skeletal and one cardiac TnI forms(stnI and cTnI, respectively)show 40% dissimilarity.Moreover,human cardiac TnI has 31 additional residues on its N-terminal end that are not present in skeletal forms, thus providing a high potential for obtaining cardiac-specific antibodies. The molecular weight of this protein is 29 kDa and therefore,it was released reasonably rapidly after acute myocardial infarction(AMI). Concentration of cTnI is very low in the normal human serum (between 0 to 3.0μg/L)but there is large amount of cTnI in myocardium, which is many times greater than the concentration of CKMB. Therefore, the concentration of cTnI goes up to 158 times the discriminator in response to an acute myocardial infarction. CTnI as one of the most specific and sensitive markers of AMI allows a reliable and timely diagnosis to be made, even in the case of concomitant skeletal muscle injury as occurred in polytraumatiozed or contused patients. In recent years, many experts raised Italy to replace other biochemistry target with cTnI to take the cardiac muscle cell damage the specificity designated object, our research goal was in had deputes cTnI in the foundation, with by the immunity New Zealand rabbit, the preparation specificity anti-cTnI multi-clone immune body, the application quantum spot is marked cTnI, then the discussion application quantum mark competition immunity analytic method examined cTnI.The following researches were carried out in the present paper:1 The preparation of the specific anti-cTnI polyclonal antibodyThe polyclonal antibodies against cardiac troponin I were prepared by immunized pure New Zealand rabbit with the purified cardiac troponin I—BSA and the antibody was purified by protein A affinity chromatography. The specific polyclonal antibodies against cardiac troponin I and BSA was obtained by nitrocellulose membrane filtration-affinity chromatography using skeletal muscle troponin I as ligand. The specificity of the antibodies was determined by ELISA(enzyme-linked immunosorbent assay) antibody showed no cross-reaction with BSA and skeletal muscle troponin I.2 Marks cTnI with the quantum spotUse water-soluble quantum (completes by Yang bai topic-based group), marks cTnI, under the uviol lamp the observed result, on the membrane has the position demonstration strong green fluorescence banding which the immune body draws a line, non-immune body position non-green fluorescence.Proved we successfully a quantum mark on the cTnI member, also the cTnI member which marked can still and the corresponding immune body member specificity union.3 The quantum mark competition immunity analytic method examines cTnI the discussionWe selected QDs to take the tracer, one step the response, the step was simple.The masculine membrane presents a green under the fluorescence the fluorescence banding, indicated we use the mark method, after the mark antigen can respond with the immune body, explained the QDs mark has not affected between the antigen immune body this kind of specificity union.The negative comparison does not have the fluorescence banding under the fluorescence, designs in us under the density condition, has not marked the antigen to occupy on the membrane immune body all epi-positions, QDs-cTnI has been unable with the immune body union.Used the competition suppression immunity analysis the principle for the first time to establish the quantum mark competition immunity analytic method to examine cTn I.This is we in studies one kind which the cTn I immune body and in the quantum mark method foundation founds for the first time to examine the cTn I new method, as a result of the instrumentation and the research time limit, only can realize at present the qualitative analysis.