CN111072778B - Monoclonal antibody for resisting duck skeletal muscle troponin I and application thereof - Google Patents
Monoclonal antibody for resisting duck skeletal muscle troponin I and application thereof Download PDFInfo
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Abstract
The invention relates to a monoclonal antibody for resisting duck skeletal muscle troponin I and application thereof, belonging to the technical field of immunology and the technical field of food safety analysis. The anti-monoclonal antibody is secreted and produced by a hybridoma cell strain with the preservation number of CCTCC NO: C202003, and the titer of the anti-monoclonal antibody is 1:105Subtype IgG1 with an affinity constant Ka of 5.9X 105L/mol. The monoclonal antibody can be used for preparing an enzyme linked immunosorbent assay kit and a colloidal gold test strip for detecting duck skeletal muscle troponin I in fresh meat and products thereof, so as to achieve the purpose of quickly and sensitively detecting duck meat-derived components in animal-derived food.
Description
Technical Field
The invention relates to a monoclonal antibody for resisting duck skeletal muscle troponin I, a hybridoma cell strain for generating the monoclonal antibody and application of the antibody in detection of duck skeletal muscle troponin I, and belongs to the technical fields of immunology and food safety analysis.
Background
In meat products, due to reasons of price, religion, health and the like, regulations established by many countries require food labels to truly and definitely mark meat sources and prohibit adulteration behaviors so as to protect the benefits of consumers, but phenomena of confusing meat varieties in the market are still very common, adulteration modes comprise means of adulteration, mixing, extraction, counterfeiting and the like, particularly the most common behaviors of adulteration and adulteration of beef and mutton products, and the adulteration behaviors greatly damage the benefits of the consumers. Due to the fact that poultry such as ducks and chickens are short in feeding period and low in price, meanwhile, the textures of duck meat and beef and mutton are highly similar, a large amount of duck meat, a small amount of beef and mutton fat and leftovers are mixed, and the processed duck meat can reach the 'false-to-false' degree, so that the duck meat is often used as a raw material for making fake beef and mutton. This adulteration impairs the consumer's interest and disturbs market regulations. Therefore, the establishment of a rapid identification method of duck meat-derived components is very important.
At present, methods for identifying meat products mainly comprise sensory identification, a nucleic acid amplification technology, a physicochemical method and an immunological method. The sensory method mainly identifies the meat through vision, smell, taste and touch, has strong subjectivity and cannot meet the detection requirement. The nucleic acid amplification technology has high detection result accuracy, is a mainstream method for detecting meat product adulteration internationally at present, has high requirements on operators, uses expensive instruments, is not suitable for application in basic laboratories and field rapid detection, and particularly has great uncertainty on detection of cooked meat products. The electronic nose is combined with gas chromatography-mass spectrometry to detect adulterated meat, so that certain accuracy is achieved, but various volatile substances need to be detected, and the operation process and result processing are complex. The immunological method has the characteristics of high sensitivity, good specificity, low cost, convenient operation and the like, and is suitable for screening large-batch samples, wherein an enzyme-linked immunosorbent assay and a colloidal gold test strip are the most common methods.
However, cooked meat products are generally subjected to high temperature and high pressure treatment, and many proteins are denatured in the process, so that antigenicity and water solubility are lost, and the establishment of an immunological method is difficult. In order to apply the immunological method to the detection of animal-derived components, it is necessary to find a specific thermostable protein as a marker antigen and then prepare a monoclonal antibody specific to the antigen. Troponin I in animal skeletal muscle has species specificity, and can be used as a heat-resistant type marker protein to distinguish meat species sources of different species of raw and cooked meat products. The duck skeletal muscle troponin I (skTnI) is taken as a target detection object, and the establishment of the duck meat derived component immunological detection and identification technology can be realized. Therefore, the preparation of the monoclonal antibody against duck skeletal muscle troponin I is of great significance for carrying out the immunological test and identification work of duck meat-derived components.
Disclosure of Invention
The invention aims to overcome the technical defects of the existing meat-derived detection technology and provides a monoclonal antibody against duck skeletal muscle troponin I, wherein the monoclonal antibody has specificity on duck skeletal muscle troponin I and is generated by a hybridoma cell strain skTnI-3E7 with the preservation number of CCTCC NO: C202003. Furthermore, the monoclonal antibody is applied to detecting duck skeletal muscle troponin I.
The technical problem of the invention is realized by the following technical scheme.
A monoclonal antibody against duck skeletal muscle troponin I is generated by a hybridoma cell strain with the preservation number of CCTCC NO: C202003. The hybridoma cell strain is named as a hybridoma cell strain skTnI-3E7, and is delivered to a China Center for Type Culture Collection (CCTCC) for preservation in 2019, 12 months and 10 days, wherein the preservation unit address is as follows: wuhan in China.
The monoclonal antibody for resisting duck skeletal muscle troponin I can be specifically combined with duck skeletal muscle troponin I, and the titer is 1:105Subtype IgG1 with an affinity constant Ka of 5.9X 105L/mol。
The monoclonal antibody against duck skeletal muscle troponin I is applied to preparation of a non-diagnosis-purpose detection product for detecting duck skeletal muscle troponin I in fresh meat and products thereof.
In the application, the non-diagnosis-purpose detection product is an enzyme-linked immunosorbent assay kit or a colloidal gold chromatography test strip.
Further, an enzyme linked immunosorbent assay kit for detecting duck skeletal muscle troponin I contains the monoclonal antibody for resisting duck skeletal muscle troponin I.
A colloidal gold chromatography test strip for detecting duck skeletal muscle troponin I contains the monoclonal antibody for resisting duck skeletal muscle troponin I.
A method for preparing the monoclonal antibody against duck skeletal muscle troponin I comprises the following steps:
(1) preparation of antigens
Taking duck skeletal muscle, removing fat and connective tissue, grinding, mixing, weighing 20g, adding 0.15M NaCl solution (1: 2M/V); mixing, ultrasonically extracting for 5min (100W), boiling for 20min, and centrifuging at 5000g for 20 min; removing the precipitate, taking the supernatant, and filtering to obtain the detection antigen. Treating 20g of ground skeletal muscle according to the method, centrifuging, collecting supernatant, and centrifuging at 121 deg.C under high pressure for 30min and 5000g for 30 min; filtering the supernatant with Whatman No. 1 filter paper, adding 90% ethanol (1:3.74V/V) into the filtrate, and standing for 2 hr; centrifuging the mixed solution at 7000g for 20min, and oven drying the precipitate to obtain immunogen extract. The separation gel concentration is 12 percent, the concentration gel is 5 percent, the antigen is identified by SDS-PAGE, and the gel is cut after electrophoresis to be used as the immune antigen.
(2) Preparing a duck skeletal muscle troponin I monoclonal antibody:
(a) animal immunization: selecting extracted immune antigen, immunizing female Balb/c mice of 6-8 weeks old, immunizing for 1 time at intervals of 2 weeks, after 3 times of immunization, cutting tail and taking blood to measure titer and inhibition rate, and selecting the mice with the best immune result for fusion;
(b) cell fusion: fusing splenocytes of the mouse selected in the step (a) with myeloma SP2/0 cells of the mouse, determining supernatant by an indirect ELISA method, selecting positive high holes, and subcloning the positive holes by a limiting dilution method until a hybridoma cell strain generating a single monoclonal antibody against duck skeletal muscle troponin I is established;
(c) large-scale preparation of monoclonal antibodies: selecting a female Balb/c mouse with a larger individual, preparing a large amount of ascites by an in vivo induced ascites method, purifying the ascites by caprylic acid-ammonium sulfate precipitation, dividing the ascites into small tubes, and storing the small tubes at the temperature of-20 ℃ to obtain the duck skeletal muscle troponin I resistant monoclonal antibody.
A method for identifying the characteristics of the monoclonal antibody for resisting duck skeletal muscle troponin I comprises the following steps:
(a) potency assay
Diluting a detection antigen to 5 mu g/mL by using a carbonate buffer solution with pH9.6 to coat a detection plate, diluting the purified duck skeletal muscle troponin I monoclonal antibody at a ratio of 1:2000, 1:4000, 1:8000, … … 1:1024000, adding the diluted antibody into a hole of an enzyme label plate, adding an HRP-labeled goat anti-mouse secondary antibody after reaction, and finally developing by using TMB (tetramethylbenzidine), wherein the result shows that the titer of the purified duck skeletal muscle troponin I monoclonal antibody reaches 1 when the concentration is 1 mg/mL: 105。
(b) Subtype determination
Subtype determination was carried out using a murine monoclonal antibody subtype identification kit purchased from Sigma, and the result showed that the subtype of the monoclonal antibody against duck skeletal muscle troponin I was IgG 1.
(c) Affinity assay
The affinity constant of the monoclonal antibody against duck skeletal muscle troponin I was determined by indirect ELISA, and the result showed that the affinity constant Ka was 5.9X 105L/mol。
(d) Specific assay
The cross reaction of monoclonal antibody of anti-duck skeletal muscle troponin I and skeletal muscle extracts of cattle, sheep, chicken, duck and fish is measured by indirect ELISA. The results show that the cross reaction rate of 3E7 with duck skeletal muscle extract is 100%, the cross reaction rate with bovine, ovine, chicken and porcine skeletal muscle extract is lower than 12.5%, and the specificity is better.
The monoclonal antibody for resisting duck skeletal muscle troponin I provided by the invention can be applied to detection of duck skeletal muscle troponin I in a sample, and is mainly applied to preparation of an enzyme-linked immunosorbent assay kit and colloidal gold test strip paper for duck skeletal muscle troponin I detection. The quality of the duck skeletal muscle troponin I resisting monoclonal antibody provided by the invention has strong controllability and repeatability, and experimental basis is provided for further establishment and application of a specific and sensitive immunoassay method through preliminary analysis and identification of the characteristics of the obtained antibody.
Drawings
FIG. 1 SDS-PAGE identification chart of duck skeletal muscle troponin I extracted according to the invention
FIG. 2 is a graph showing the measurement of the subtype of the monoclonal antibody against duck skeletal muscle troponin I according to the present invention
FIG. 3 is a graph showing the determination of the affinity of the monoclonal antibody against duck skeletal muscle troponin I according to the present invention
Detailed Description
The present invention is further described in detail with reference to the following specific embodiments, which are not intended to limit the scope of the present invention.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example A preparation of Duck skeletal muscle troponin I antigen of the present invention
Taking duck skeletal muscle, removing fat and connective tissue, grinding, mixing, weighing 20g, adding 0.15M NaCl solution (1: 2M/V); mixing, ultrasonically extracting for 5min (100W), boiling for 20min, and centrifuging at 5000g for 20 min; removing the precipitate, taking the supernatant, and filtering to obtain the detection antigen. Treating 20g of ground skeletal muscle according to the method, centrifuging, collecting supernatant, and centrifuging at 121 deg.C under high pressure for 30min and 5000g for 30 min; filtering the supernatant with Whatman No. 1 filter paper, adding 90% ethanol (1:3.74V/V) into the filtrate, and standing for 2 hr; centrifuging the mixed solution at 7000g for 20min, and oven drying the precipitate to obtain immunogen extract. The separation gel concentration was 12% and the concentration gel was 5% and the antigen was identified by SDS-PAGE, as shown in FIG. 1, which shows 2 bands with clear bands and darker colors, corresponding to molecular weights of about 37kDa and 24kDa, corresponding to T, I subunits of skeletal muscle troponin reported in the literature. A24 kDa band cut gel was selected as the immunizing antigen.
EXAMPLE two preparation of monoclonal antibody against duck skeletal muscle troponin I according to the present invention
(1) Animal immunization: immunizing female Balb/c mice of 6-8 weeks old with the prepared immune antigen for 1 time at intervals of 2 weeks, wherein the immune process is shown in Table 1, cutting off the tail after three-immunization, taking blood, measuring titer and inhibition rate, and selecting the mice with the best immune result for fusion;
TABLE 1 immunization protocol
(2) Cell fusion: fused mice are bled by eyes, serum is used as positive control, spleens are taken out under aseptic condition after cervical removal and sacrifice, splenocytes are prepared and fused with SP2/0 cells according to the proportion of 5:1 by PEG, fused cell suspension is added into a 96-well plate which is paved with feeder cells, the 96-well plate is placed at 37 ℃ and 5% CO2Culturing in an incubator;
(3) screening positive hybridoma cell strains: the fused cells were checked for contamination the next day, and were replaced with HT medium 10 days after fusion. And (3) after liquid replacement, screening positive holes by using indirect ELISA (enzyme-linked immunosorbent assay), selecting holes with strong positive, single clone and good cell state as much as possible, carrying out subcloning by using a limiting dilution method, and simultaneously carrying out amplification culture and cryopreservation until a monoclonal cell strain with single secretory antibody is established.
(4) Large-scale preparation of monoclonal antibodies: adopting a method of inducing ascites in mice, taking healthy Balb/c female mice, injecting 0.5mL of paraffin oil into each mouse, adjusting 10 g of clone positive hybridoma after 7 days6and/mL, injecting 1mL into the abdominal cavity of each mouse, taking ascites after 7-9 days, centrifuging, removing fat, purifying by using caprylic acid-ammonium sulfate, freeze-drying, and storing at-20 ℃ to obtain the duck skeletal muscle troponin I resistant monoclonal antibody.
EXAMPLE III characterization of monoclonal antibodies against duck skeletal muscle troponin I according to the invention
(1) The titer measurement adopts an indirect ELISA method, and comprises the following specific steps:
coating: diluting the coated antigen with carbonate buffer solution until the concentration is 5 mug/mL, and keeping the concentration of the coated antigen in a 96-well enzyme label plate at 100 mug/well for overnight at 4 ℃;
washing: recovering the coated plate to room temperature, decanting off coating solution, adding 300 μ L of washing solution into each well, standing for L min each time, washing for 3 times, and patting to dry for the last time;
and (3) sealing: adding 200 μ L of lotion containing 10% calf serum into each well, and keeping the temperature at 37 deg.C for 1 h; pouring off the confining liquid, washing for 3 times, and patting dry;
adding a primary antibody: diluting the monoclonal antibody with washing solution at a ratio of 1:2000 times, adding 100 μ L per well, setting blank control well (PBS) and negative control (negative serum), and standing at 37 deg.C for 45 min; pouring out primary antibody, washing for 3 times, and patting dry;
adding an enzyme-labeled secondary antibody: adding 100 μ L of HRP enzyme-labeled goat anti-mouse IgG diluted by 10000 times into each well, and standing at 37 deg.C for 30 min; pouring out the secondary antibody, washing for 3 times, and patting dry; color development: adding 100 mu L of substrate developing solution into each hole, and reacting for 15min at 37 ℃ in a dark place;
and (4) terminating: adding 50 mu L of stop solution into each hole to stop the reaction;
and (3) detection: the absorbance value at a wavelength of 450nm was measured (A450 nm).
And (4) judging a result: and (A sample well-A blank)/(A negative control-A blank) is not less than 2.1, and when A450nm of the negative control is less than 0.2, the dilution factor of the antibody is the antibody titer.
Through determination, the titer of the monoclonal antibody 3E7 for resisting duck skeletal muscle troponin I at the concentration of 1mg/ml reaches 1:105。
(2) Subtype determination
Subtype determination was carried out using a murine monoclonal antibody subtype identification kit purchased from Sigma. The results are shown in fig. 2, the monoclonal antibody secreted by the hybridoma cell has obvious difference with the secondary antibody of different subclasses, the value of A450nm of the 3E7 and the IgG1 secondary antibody is the highest, the color with the IgM secondary antibody is weak, the color with the IgG2a, the IgG2b, the IgG3 and the IgA secondary antibody is hardly developed, and the antibody type secreted by the hybridoma cell 3E7 is Ig G1.
(3) Affinity assay
The affinity constant (Ka) was determined by a non-competitive ELISA method. The method comprises the following specific steps:
coating: diluting the coated antigen with carbonate buffer solution until the concentration is 1, 0.5, 0.1 and 0.05 mu g/mL, respectively coating the 96-well ELISA plate with 100 mu L/well, and standing overnight at 4 ℃; pouring out the coating solution, washing for 3 times, and patting dry;
and (3) sealing: adding 200 μ L of lotion containing 10% calf serum into each well, and keeping the temperature at 37 deg.C for 1 h; pouring off the confining liquid, washing for 3 times, and patting dry;
adding a monoclonal antibody: diluting the monoclonal antibody with washing solution at a ratio of 100 μ g/mL, adding 100 μ L per well, and keeping the temperature and humidity at 37 deg.C for 45 min; pouring out primary antibody, washing for 3 times, and patting dry;
adding an enzyme-labeled secondary antibody: adding 100uL of goat anti-mouse IgG labeled with HRP enzyme and diluted by 10000 times into each hole, and standing at 37 ℃ for 30 min; pouring out the secondary antibody, washing for 3 times, and patting dry;
color development: adding 100 mu L of substrate developing solution into each hole, and reacting for 15min at 37 ℃ in a dark place;
and (4) terminating: adding 50 mu L of stop solution into each hole to stop the reaction;
and (3) detection: the absorbance value at a wavelength of 450nm was measured (A450 nm).
The logarithm of the antibody concentration is used as the abscissa, the OD value is used as the ordinate, an S-shaped curve is drawn, the S-shaped curve is shown in figure 3, and the affinity constant Ka of the monoclonal antibody against duck skeletal muscle troponin I is calculated to be 5.9 multiplied by 105L/mol。
(4) Specific assay
The indirect competitive ELISA method is adopted, and the specific steps are as follows:
coating: diluting the coating antigen (bovine, sheep, chicken, duck, fish skeletal muscle troponin I extract) with carbonate buffer solution to the concentration of 5 μ g/mL, and standing overnight at 4 deg.C with a 96-well ELISA plate of 100 μ L/well; pouring out the coating solution, washing for 3 times, and patting dry;
and (3) sealing: adding 200 μ L of lotion containing 10% calf serum into each well, and keeping the temperature at 37 deg.C for 1 h; pouring off the confining liquid, washing for 3 times, and patting dry;
adding a primary antibody: diluting the monoclonal antibody with washing solution at a ratio of 1:2000 times, adding 100 μ L per well, setting blank control well (PBS) and negative control (negative serum), and standing at 37 deg.C for 45 min; pouring out primary antibody, washing for 3 times, and patting dry;
adding an enzyme-labeled secondary antibody: adding 100 μ L of HRP enzyme-labeled goat anti-mouse IgG diluted by 10000 times into each well, and standing at 37 deg.C for 30 min; pouring out the secondary antibody, washing for 3 times, and patting dry;
color development: adding 100 mu L of substrate developing solution into each hole, and reacting for 15min at 37 ℃ in a dark place;
and (4) terminating: adding 50 mu L of stop solution into each hole to stop the reaction;
and (3) detection: the absorbance value at a wavelength of 450nm was measured (A450 nm).
And (4) judging a result: when the volume of the sample well A-A blank)/(the negative control A-A blank) is not less than 2.1 and the volume of the negative control A450nm is less than 0.2, the dilution ratio of the antibody is the antibody titer.
TABLE 2 Cross-reaction results
The results are shown in Table 2, 3E7 has a cross reaction rate of 100% with duck skeletal muscle extract, a cross reaction rate of less than 12.5% with skeletal muscle extract of cattle, sheep, chicken and pig, and good specificity.
Example four applications of the monoclonal antibody against duck skeletal muscle troponin I of the present invention
The embodiment is an application example of the duck skeletal muscle troponin I resisting monoclonal antibody disclosed by the invention after an enzyme-linked immunosorbent assay kit or a colloidal gold chromatography test strip method for detecting duck skeletal muscle troponin I is established, and the monoclonal antibody can be used for detecting duck skeletal muscle troponin I in fresh meat and products thereof.
(1) Sample pretreatment
Removing fat and connective tissue from fresh meat, processing meat food, removing casing, weighing 1g, adding 0.15M NaCl solution 2ml (1:2w/v), homogenizing, heating with boiling water for 20min, centrifuging at 2000g for 30 min; the precipitate was removed, the supernatant was filtered through Whatman No. 1 filter paper, and the filtrate was collected for detection.
(2) The antibody is applied to the detection of an enzyme-linked immunoassay kit method
The detection principle of the kit is a double-antibody sandwich ELISA method. Coating a micropore lath with an antibody (3E7) to prepare a solid-phase capture antibody, sequentially adding a standard substance (sample) and a polyclonal antibody into micropores coated with a monoclonal antibody, combining with a goat anti-rabbit antibody marked by horseradish peroxidase to form an antibody-antigen-antibody-enzyme-labeled antibody compound, adding TMB for color development, wherein the compound is blue and finally yellow under the action of acid, and the color depth is in positive correlation with the content of duck skeletal muscle troponin I in the sample.
And (3) detection: diluting duck skeletal muscle troponin I monoclonal antibody with carbonate buffer solution at a ratio of 1:500, coating at 100 μ L/well, and standing overnight at 4 deg.C; washing and patting to dry; blocking with 1% gelatin in PBS, 200. mu.L/well, incubating at 37 ℃ for 2 h; washing and drying; adding a sample to be detected, incubating for 30min at 37 ℃ in a hole of 100 mu L, washing, and patting dry; adding polyclonal antibody diluted at a ratio of 1:2000, 100 μ L/well, incubating at 37 deg.C for 30min, washing, and patting to dry; adding diluted horse radish peroxidase-labeled goat anti-rabbit antibody, incubating at 37 deg.C for 30min, washing, and patting to dry; adding 100 μ L/well of color developing solution, developing for 15min, adding 50 μ L/well of stop solution, and measuring.
And (4) judging a result:
(a) quantitative analysis: respectively calculating the average absorbance values of the standard substance and the sample to be detected, wherein the absorbance value (B) of the standard substance or the sample to be detected is a vertical coordinate, the addition concentration is a horizontal coordinate, and drawing a standard curve; substituting the absorbance value of the sample to be detected into the standard curve to obtain the corresponding concentration, and multiplying the concentration by the dilution factor to obtain the content of the sample.
(b) And (3) qualitative analysis: and comparing the average absorbance value of the sample to be detected with the absorbance value of the standard substance to obtain the concentration range of the sample to be detected.
(3) Method for detecting colloidal gold chromatography test paper strip by using antibody
The reaction principle adopts a double-antibody sandwich method to qualitatively detect duck skeletal muscle troponin I, duck skeletal muscle troponin I existing in a sample is firstly combined with a gold particle-labeled antibody in the process of moving up along a test strip to form a gold-labeled antibody-duck skeletal muscle troponin I compound, a capture antibody fixed on an NC membrane is combined with duck skeletal muscle troponin I in the compound, and the color development intensity of a T position (a detection line) is in direct proportion to the content of the duck skeletal muscle troponin I in the sample.
And (3) detection: taking out the duck skeletal muscle troponin I colloidal gold chromatography test strip, inserting the sample end into the sample liquid to be detected, wherein the insertion depth is not more than the mark line, taking out the detection test strip in about 10-20 seconds, horizontally placing, observing for 3-5 minutes to judge the detection result, and after 10 minutes, the result is invalid.
And (4) judging a result: a brownish red line is displayed at the position (quality control line) of a corresponding quality control area C on the envelope film, and a brownish red line is not displayed at the position T of the detection area, which indicates that the detection result is negative, and indicates that the sample to be detected does not contain duck skeletal muscle troponin I; two brown-red lines are displayed at the T, C position on the envelope membrane, the result is positive, and the result indicates that the sample to be detected contains duck skeletal muscle troponin I; when the quality control zone C does not show a brownish red strip, the test paper is judged to be invalid no matter whether the detection zone T shows a brownish red strip or not.
The above-described embodiments are intended only to illustrate the technical idea and advantages of the invention, and the invention may be subject to other variants, as will be known to those skilled in the art, which are merely exemplary of the scope of protection of the invention described above, and many routine modifications and other embodiments are possible for those skilled in the art within the scope of protection defined by the invention, which are all intended to be covered by the scope of protection of the invention.
Claims (6)
1. A monoclonal antibody against duck skeletal muscle troponin I is characterized in that the monoclonal antibody is generated by a hybridoma cell strain skTnI-3E7 with the preservation number of CCTCC NO: C202003.
2. A hybridoma cell strain skTnI-3E7 capable of producing the monoclonal antibody against duck skeletal muscle troponin I as claimed in claim 1, which is preserved in China center for type culture Collection with the preservation number of CCTCC NO: C202003, and the preservation date is 2019, 12 months and 10 days.
3. Use of the monoclonal antibody against duck skeletal muscle troponin I according to claim 1 for the preparation of a test product for the non-diagnostic purpose of detecting duck skeletal muscle protein I in a biological sample.
4. The use of claim 3, wherein the non-diagnostic detection product is an enzyme linked immunosorbent assay kit or a colloidal gold chromatography strip.
5. An enzyme linked immunosorbent assay kit for detecting duck skeletal muscle troponin I, which is characterized by comprising the monoclonal antibody against duck skeletal muscle troponin I as claimed in claim 1.
6. A colloidal gold chromatography test strip for detecting duck skeletal muscle troponin I, which comprises the monoclonal antibody against duck skeletal muscle troponin I according to claim 1.
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| CN102030825A (en) * | 2010-10-22 | 2011-04-27 | 上海贝西生物科技有限公司 | Troponin I resisting monoclonal antibody and application thereof |
| CN109709338A (en) * | 2018-12-28 | 2019-05-03 | 河北省科学院生物研究所 | Detect ox or the enzyme linked immunological kit of ovine skeletal muscle Troponin I and the preparation method and application thereof |
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| CN102030825A (en) * | 2010-10-22 | 2011-04-27 | 上海贝西生物科技有限公司 | Troponin I resisting monoclonal antibody and application thereof |
| CN109709338A (en) * | 2018-12-28 | 2019-05-03 | 河北省科学院生物研究所 | Detect ox or the enzyme linked immunological kit of ovine skeletal muscle Troponin I and the preparation method and application thereof |
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| 猪骨骼肌肌钙蛋白Ⅰ基因的表达及其单克隆抗体的制备;孟宪荣等;《中国畜牧兽医学会2009学术年会论文集（上册）》;20091231;第312-316页 * |
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