CN110850079A - Application of diagnosis marker APOA1 for effect evaluation after liver transplantation - Google Patents
Application of diagnosis marker APOA1 for effect evaluation after liver transplantation Download PDFInfo
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Abstract
The invention discloses application of a serum polypeptide molecular diagnostic marker APOA1 for evaluating the effect after liver transplantation, wherein the amino acid sequence of the serum polypeptide molecular diagnostic marker APOA1 is shown in SEQ ID No. 1. This molecule is called APOA1, APOA1 is apolipoprotein a1, with an exact molecular weight of 2997.64 daltons. The APOA1 has better recovery after liver transplantation than that of the serum of a patient before operation and shows specific high expression, and the expression level of APOA1 detected by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) or APOA1 detected by ELISA method can be used as a detection method for evaluating the serum after liver transplantation.
Description
Technical Field
The invention belongs to the technical field of biomarkers, and relates to application of a diagnosis marker APOA1 for effect evaluation after liver transplantation.
Background
Since the first application of Liver Transplantation (LT) technology to clinic by Starzl in 1963, liver transplantation has become the only effective means for treating end-stage liver diseases at present after half a century of development. However, the occurrence of various complications after liver transplantation, such as infection, acute rejection, liver failure, etc., presents a significant challenge to clinical work. How to early identify and timely intervene in postoperative complications is a key to improving survival time of transplant patients.
Due to the extensive liver function, in the whole treatment process of liver transplantation, the pathophysiological changes of all systems of the whole body of a patient are complex, and factors influencing the outcome of diseases are various. Liver biopsy is considered the gold standard for diagnosing postoperative complications in this type of patient, and has its clinical limitations as an invasive test. After the liver transplantation, the liver function condition of the patient after transplantation is observed by various enzymology and bilirubin indexes, and the prognosis of the patient is predicted. Therefore, the laboratory examination is low in price, simple and easy to implement, and is often used as a conventional index for postoperative reexamination of liver transplantation patients clinically. However, the changes and specific action mechanism of liver function index after liver transplantation are not completely clear, and many influencing factors exist. In recent years, MELD score is clinically used more, and blood lactate level is used as a tool for evaluating prognosis of liver transplantation patients, but the prediction accuracy is still controversial. Therefore, it is very important for the development of the patient's condition to make timely assessment on the outcome of the patient's condition before and after surgery in order to achieve early intervention treatment, and there are few reports on the predictor for assessing the development of the liver transplantation patient's condition in clinical work. The research and discovery of new post-transplantation monitoring of the liver function recovery and prognosis index of patients have important clinical significance.
Before any pathological change occurs in any disease, the intracellular proteins are altered in composition and quantity and are reflected by the pattern of proteins in the serum. Therefore, by comparing the expression of different proteins in the serum of different disease populations, it is possible to screen out disease-related marker molecules. Serum proteomics refers to the study of all proteins expressed in the serum of a selected target population, and on the basis of establishing a normal Protein Expression Map (PEM), differential protein spots of the proteins are searched, and disease-related proteins are identified, so that the structure and function of the proteins are further studied, and a new way is developed for studying major disease pathophysiological mechanisms, specific markers for early diagnosis, drug action targets and the like. A large amount of proteins and polypeptides exist in human serum, and the existence, deletion and expression of partial proteins and polypeptides are closely related to the health degree of human beings, so that the human serum becomes a biomarker for disease diagnosis.
Serum diagnosis is considered to be the most recent and effective method for early diagnosis of cancer. The method judges the occurrence and development of the tumor by searching tumor markers in blood, particularly protein markers in the blood, thereby realizing early diagnosis of the tumor. A large amount of proteins and polypeptides exist in human serum, and the existence, deletion and expression of partial proteins and polypeptides are closely related to the health degree of human beings, so that the human serum becomes a biomarker for disease diagnosis.
At present, few reports are available on the prediction molecules for the effect evaluation after liver transplantation.
Disclosure of Invention
The invention aims to provide application of a diagnostic marker APOA1 for evaluating the effect after liver transplantation.
In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:
the invention discloses an application of a serum polypeptide molecular diagnostic marker APOA1 in preparation of a reagent for evaluating and diagnosing the effect of a liver transplantation, wherein the amino acid sequence of the serum polypeptide molecular diagnostic marker APOA1 is shown in SEQ ID No. 1.
Preferably, the serum polypeptide molecular diagnostic marker APOA1 is apolipoprotein A1, and the molecular weight is 2997.64 daltons.
Preferably, the detection parameter of the serum polypeptide molecular diagnostic marker APOA1 in serum is 8.99-13.07 ug/mL.
Preferably, the reagent for evaluating the effect after liver transplantation is a serum polypeptide molecular reagent for ELISA detection of evaluation of the effect after liver transplantation.
The invention also discloses application of the molecule combined with the serum polypeptide molecular diagnostic marker APOA1 in preparation of a diagnostic reagent for evaluating the effect after liver transplantation.
The invention also discloses application of the molecule combined with the APOA1 protein in preparing a diagnostic reagent for evaluating the effect after liver transplantation.
Compared with the prior art, the invention has the following beneficial effects:
the invention discloses a serum polypeptide molecule for evaluating the effect after liver transplantation, the amino acid sequence of the serum polypeptide molecule is shown in SEQ.ID.NO.1, and the molecule is called APOA 1. APOA1 is apolipoprotein a1, with an exact molecular weight of 2997.64 daltons. The APOA1 has obviously higher expression in serum detection of patients with good recovery after liver transplantation than before operation, and the expression range in serum of normal healthy people is as follows: 20.19-12.77 ug/mL; expression ranges in serum of patients before liver transplantation were: 4.74-11.7 ug/mL; the expression ranges in serum of patients recovering well after liver transplantation are: 7.08-15.13ug/mL, and there was a very significant difference in expression between the different groups (P < 0.001).
Since APOA1 is expressed in blood serum of patients with good recovery after liver transplantation and is lower than that of healthy normal people, APOA1 can be used as a serum diagnostic marker for evaluating the effect after liver transplantation; the expression level of APOA1 or APOA1 is detected by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and the method can be used as a method for detecting patients with good recovery after liver transplantation. And aiming at ELISA detection of good recovery of serum diagnosis of patients after liver transplantation, APOA1 can be used as a new target point of ELISA detection drugs.
Drawings
FIG. 1 is a diagram showing the results of Flex analysis of specifically low-expressed 7 protein polypeptides in serum of a patient before liver transplantation: through the expression comparison of M/Z:2997.64 in a patient before liver transplantation (red), a patient with good recovery after liver transplantation (green) and a normal healthy population (blue), the protein polypeptide peak pattern of M/Z:2997.64 is found to be remarkably low expressed in the serum of the patient before liver transplantation, so that the sequence identification is carried out on the protein polypeptide peak pattern and the first-choice further identification as a marker is carried out.
FIG. 2 shows the results of gel chromatography of serum protein polypeptides from patients before liver transplantation: the abscissa in the chromatogram represents the sample outflow time, the ordinate represents the relative abundance of the polypeptide, the set time of the chromatogram is 78min, fractions are collected from 5min, the polypeptide components are mainly separated after 6min, a gradient elution mode is adopted, the elution efficiency is improved, and the capture time is set to collect the fractions: 25 peptide fractions were collected and the peak for the 2.9K Da peptide of interest flowed out at about 6 min.
FIG. 3 is a MS/MS mass spectrometric identification profile of APOA 1;
fig. 4 shows the expression levels of APOA1 protein in different groups: the results of enzyme-linked immunosorbent assay analysis show that the expression level of APOA1 in different detection groups is normal healthy people, patients with good recovery after liver transplantation, and patients before liver transplantation, and the three groups have significant difference.
Detailed Description
In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It is noted that the terms "comprises" and "comprising," and any variations thereof, as used in the description and claims of this invention, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
The invention is described in further detail below with reference to the accompanying drawings:
the serum polypeptide molecule for evaluating the effect after liver transplantation is a newly screened serum diagnosis marker for evaluating the effect after liver transplantation, has specificity in expression, and can be applied to diagnosis of evaluating the effect after liver transplantation.
The specific screening of the serum diagnosis marker for the effect evaluation after the liver transplantation comprises the following steps:
the method comprises the steps of firstly, separating and extracting serum protein polypeptides of good postoperative recovery and normal healthy people before liver transplantation by using a liquid protein chip technology, capturing serum protein polypeptide maps of the good postoperative recovery and normal healthy people before liver transplantation by using a matrix-assisted laser desorption ionization time-of-flight mass spectrometry technology, comparing and analyzing the difference of serum protein polypeptide spectrogram of the good postoperative recovery and normal healthy people before liver transplantation by using ClinProTools2.1 software, finding out protein polypeptide scores with significant differential expression among groups, and screening out serum markers for effect evaluation after liver transplantation from protein polypeptide peaks with significant high expression in serum of patients with good postoperative recovery.
The verification of the screened polypeptide serum diagnostic marker for the post-liver transplantation effect evaluation is as follows:
the protein polypeptide mixture separated from the serum of a patient with good recovery after liver transplantation is divided into 20-30 components by HPLC, secondary mass spectrum identification is carried out on the protein polypeptide mixture, an enzyme-linked immunosorbent assay is carried out on the identified protein polypeptide, and serum regression verification results prove that the protein polypeptide mixture is remarkably high in expression in the serum of the patient with good recovery after liver transplantation compared with the serum of the patient before operation, has specificity and can be used as a biomarker for serum screening of effect evaluation after liver transplantation.
1. Collecting and processing a sample:
10 (7 males; 3 females; mean age 51 years) patients undergoing classical orthotopic liver transplantation and 10 normal healthy populations (7 males; 3 females; mean age 50 years) collected from hepatobiliary surgery (2016. 3.2016. to 2016. 12. months) at the first subsidiary hospital of the university of transport, Xian. The sample considers factors such as age, sex, collection time, whether the storage conditions are consistent, whether basic diseases exist and the like. Collecting blood of the collected person with fasting state, collecting 5mL whole blood with vacuum blood collection tube (yellow cap, with isolation gel), and standing at room temperature for 30 min; centrifuging at room temperature for 5min (3000g), subpackaging the upper layer serum into 100 μ L/tube, immediately storing at-80 deg.C, and avoiding repeated freeze thawing.
Reagents and instrumentation:
the serum proteins were extracted using the magnetic bead kit "weak cationic" (MB-WCX) from Bruker, Germany, and spectrally pure (HPLC grade) acetonitrile, trifluoroacetic acid (Merck, Germany), α -cyano-4-hydroxycinnamic acid (HCCA) (Sigma, USA).
Magnetic bead separator, 600/384Anchor chip target plate and AutoFlexIII Matrix Assisted Laser Desorption Ionization Time of Flight mass spectrometry (Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry, MALDI-TOF-MS) (Bruker Daltonics, Germany).
2. Preparation of serum protein samples
Capturing serum protein polypeptide by weak cation (MB-WCX) magnetic beads, which comprises the following specific operation steps:
① mixing the magnetic bead suspension completely for 1 min;
② adding 10 μ L MB-WCX binding solution and 10 μ L MB-WCX magnetic bead into PCR tube, mixing, adding 5 μ L serum, mixing for at least 5 times, and standing for 5 min;
③ placing PCR tube into magnetic column separator to make magnetic beads adhere to wall for 1min, and removing supernatant after the liquid is clear;
④ adding 100 mu LMB-WCX washing liquid, moving the PCR tube on a magnetic column separator back and forth for 10 times, and removing the supernatant after the magnetic beads adhere to the wall;
⑤ repeating steps ③ and ④ twice, then adding 5 mu of LMB-WCX eluent to wash adherent magnetic beads, repeatedly blowing for 10 times, allowing the magnetic beads to adhere for 2min, and transferring the supernatant into a clean centrifuge tube;
⑥ adding 5 μ LMB-WCX stable solution into centrifuge tube and mixing, the extracted protein polypeptide can be used for direct MALDI-TOF-MS detection or frozen in refrigerator at-20 deg.C for 24h mass spectrum analysis.
Mass spectrometry analysis:
mixing 1 μ L of protein sample obtained by separation and collection with 10 μ L of substrate α -cyano-4-hydroxycinnamic acid, placing 1 μ L of the mixture on an Anchorchip target plate (Bruker, Germany), placing each sample with three targets for three times, drying at room temperature, putting the target plate into a mass spectrometer for flight time mass spectrometry, correcting standard products by FlexControl 2.0 software, starting sample detection, generating a mass spectrogram after each sample is subjected to laser targeting for 300 times (5 times of point targeting and 2 × 30 times of targeting each time), obtaining a protein polypeptide spectrogram consisting of different mass-to-nuclear ratios (m/z), analyzing protein polypeptide spectrums of two groups of serum samples by ClinProTools2.1 software in combination with a genetic algorithm and other biological statistics and bioinformatics methods, carrying out normalization smoothing treatment on the total ion flow chart, eliminating chemical and electrical physical noise, analyzing proteins with difference between groups, calculating the difference size, arranging the proteins from large to small according to find out protein peak expression with difference (P < 0.001).
The method comprises the steps of processing serum samples of good postoperative recovery and normal healthy population before liver transplantation by using a magnetic bead separation system, carrying out MALDI-TOF-MS analysis, drawing protein polypeptide maps of each sample of serum of good postoperative recovery and normal healthy population before liver transplantation, detecting 61 protein polypeptide peak maps in a molecular weight range of 1000 Da-10000 Da, and enabling the three-time repeated stability of each sample to be high.
Analyzing serum protein polypeptide maps of patients with good recovery before and after liver transplantation captured by mass spectrometry by using ClinProTools2.1 software, comparing and analyzing the serum protein polypeptides of the patients with good recovery after the liver transplantation, and detecting 7 protein polypeptide peak maps (P <0.05) with significant differences in total, wherein the 7 protein polypeptide peak maps have very significant differences among three groups (P <0.001), wherein 5 protein polypeptides are significantly up-regulated in the patients with good recovery after the liver transplantation, and the expression of the rest 2 protein polypeptides is significantly down-regulated in the patients with good recovery after the liver transplantation, which is specifically shown in Table 1:
TABLE 1
When Flex analysis software analysis is carried out on 5 specific protein polypeptides with high expression in the serum of the patient with good postoperative recovery in the table 1, the result is shown in the figure 1, and through expression comparison of M/Z:2997.87 in the patients with good postoperative recovery (red) and the patients with good postoperative recovery (green) and the normal healthy population (blue), the protein polypeptide peak diagram of M/Z:2997.87 is found to be significantly high expressed in the serum of the patient with good postoperative recovery compared with the serum of the patient before operation, so that the sequence identification is carried out on the protein polypeptide peak diagram and the first-choice further identification is carried out as a marker.
3. Sequence identification of serum potential marker for evaluating effect after liver transplantation
Specifically, a liquid chromatography separation and mass spectrometry combined technology is adopted to identify a serum polypeptide marker M/Z:2997.87 of a patient with good recovery after liver transplantation, two-dimensional gel chromatography separation is carried out on the residual serum protein polypeptide after the magnetic bead separation and collection of mass spectrometry by adopting Nano acquisition UPLC of Waters company, and 25 peptide fragment fractions are collected: the 2.9kDa peak was shed at about 6min and detected in the collection; and then, the sequence identification is carried out on the protein polypeptide M/Z:2997.87 with the expression up-regulated in the serum of a patient with good recovery after liver transplantation by using an LTQ Orbitrap XL mass spectrum system of Thermo Fisher company.
3.1 sample Pre-treatment
Mixing the extracted protein samples, converting the protein samples to 13300 revolutions for 5 minutes, taking supernatant, drying the supernatant by a freeze dryer to ensure that the final volume is 50ul to obtain liquid A, extracting the liquid A by an Agilent zip extraction column, and concentrating the liquid A. the treatment method comprises the steps of blowing and beating ① zip columns for 5 times by 100% acetonitrile to activate the columns, repeatedly blowing and sucking ② activated zip columns in the liquid 1 for 10 times to avoid bubbles as much as possible, washing 3 times of the zip columns by ③ 40% ACN 0.2% TFA aqueous solution, repeatedly blowing and sucking ④ and zip columns in 0.2% TFA to elute a sample eluent, repeating the steps 1-4 and 30 times by ⑤, merging 30 times of eluent 2 by ⑥, freezing and drying the eluent to 10ul for mass spectrum identification.
3.2 chromatographic separation
The original sample was added with 10ul of mobile phase A and transferred to a sample vial for a total of 20 ul.
One-dimensional ultra-high performance liquid phase system: nano Aquity UPLC (Waters Corporation, Milford, USA). A chromatographic column:
a trapping column:
C18,3μm,0.10×20mm,nanoAcquityTMColumn
and (3) analyzing the column:
C18,1.9μm，0.15×120mm,nanoAcquityTMColumn
mobile phase A: 5% acetonitrile, 0.1% formic acid in water
The mobile phase B is 95 percent of acetonitrile and 0.1 percent of aqueous solution of formic acid; all solutions were HPLC grade.
The collecting flow rate is 600nl/min, the collecting time is 3min, and the analysis flow rate is 400 nl/min; the analysis time is 60min, and the temperature of a chromatographic column is 35 ℃; the Partial Loop mode was injected in a volume of 18. mu.l.
The gradient elution procedure is shown in table 2 below:
TABLE 2
| Time (min) | A | B | Flow rate (nL/min) |
| 0 | 95％ | 5％ | 600 |
| 16 | 90％ | 10％ | 600 |
| 51 | 78％ | 22％ | 600 |
| 71 | 70％ | 30％ | 600 |
| 72 | 5％ | 95％ | 600 |
| 78 | 5％ | 95％ | 600 |
The results of gel chromatography are shown in FIG. 2. The abscissa in the chromatogram represents the sample outflow time, the ordinate represents the relative abundance of the polypeptide, the set time of the chromatogram is 78min, fractions are collected from 5min, the polypeptide components are mainly separated after 6min, a gradient elution mode is adopted, the elution efficiency is improved, and the capture time is set to collect the fractions: 25 peptide fractions were collected and the peak for the 2.9K Da peptide of interest flowed out at about 6 min.
3.3LTQ-Orbitrap XL Mass Spectrometry
Q-exact HF mass spectrometry system (Thermo Fisher Co., Ltd.) was used. Nano liter flow rate HPLC liquid phase system Easy nLC 1000, spray voltage 1.8 kV; the mass spectrum scanning time is 78 min; the experimental modes are data dependence (DataDependency) and Dynamic Exclusion (Dynamic Exclusion), and parent ions are added into an Exclusion list for 80 seconds after being cascaded for 2 times within 60 seconds; the scanning range is 300-1400 m/z; the first-order scan (MS) uses obirrap with a resolution set to 70000; CID and secondary scanning use LTQ; selecting single isotope of 20 ions with the strongest intensity as parent ion in MS spectrogram to carry out MS/MS (single charge exclusion without being used as parent ion).
And (3) data analysis: database searching identification is carried out by using data analysis software mascot incorporation PD 2.0. The error of parent ion is set to be +/-15 ppm, the error of fragment ion is set to be 20mmu, the enzyme cutting mode is non-enzyme cutting, and the variable modification is M (Methionine) methionine oxidation. The search result parameter is set to be equal to or less than 0.01.
2997.87 for m/z; uniprot: P02647; gene Symbol ═ APOA 1; the sequence is as follows: llpvlesfskvslsleeytknttq.
2997.87 is called APOA1, APOA1 is apolipoprotein A1, its precise molecular weight is 2997.64 dalton, and its amino acid sequence is: llpvlesfskvslsleeytklntq. - (as shown in seq. id. No. 1).
Therefore, the method for detecting the expression level of APOA1 by using a matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS) or detecting the expression level of APOA1 by using an ELISA method can be used as a method for evaluating and detecting the effect after liver transplantation.
APOA1 was suggested to be a protein specifically associated with good recovery after liver transplantation and was further verified by ELISA assay.
4. ELISA serum validation assay for good recovery of serum APOA1 expression in patients after liver transplantation
1) Serum samples: serum verification analysis by ELISA was performed by collecting 10 cases of serum from patients before liver transplantation (8 cases for male, 2 cases for female; mean age 52), 10 cases of serum from matched patients with good postoperative recovery (8 cases for male, 2 cases for female; mean age 52), and 10 cases of serum from normal healthy population (7 cases for male; 3 cases for female; mean age 53.5). All serum samples come from Xian traffic university's first subsidiary college, and the collection time is 1 month in 2017 to 9 months in 2017;
2) the detection method comprises the following steps: the expression level of serum APOA1 of patients with good recovery before and after liver transplantation and normal healthy people is detected by an enzyme-linked immunosorbent assay (ELISA), and the kit is purchased from American R & D company. The kit adopts a double-antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA): to the coated microwells previously coated with anti-human APOA1 protein (APOA1) antibody, the specimen, the standard, and the HRP-labeled detection antibody were added in this order, incubated, and washed thoroughly. The color is developed with the substrate TMB, which is converted to blue by the catalysis of peroxidase and to the final yellow color by the action of an acid. The shade of the color was positively correlated with the APOA1 protein (APOA1) in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the sample concentration was calculated. The specific experimental steps refer to the kit specification, and the positive judgment standard is defined according to the kit specification;
3) the statistical method comprises the following steps: one-way analysis of variance (ANOVA) and T-test of independent samples were performed using graphpad. prism. v5.01 software;
4) and (4) analyzing results: and (4) analyzing results: the results of analysis by enzyme linked immunosorbent assay showed that the expression level of APOA1 in different detection groups was normal healthy population > good postoperative recovery patient > patient before liver transplantation, and the specific results are shown in table 3 and fig. 4, with significant differences between each two of the three groups.
TABLE 3
The result of ELISA detection of APOA1 in serum of patients with good recovery before and after liver transplantation and normal healthy people shows that the expression of APOA1 has specificity: the range of expression in serum in patients prior to liver transplantation was: 4.74-11.7 ug/mL; the range of expression in sera of patients with good postoperative recovery was: 7.08-15.13 ug/mL; the expression range in the serum of patients with normal healthy population is as follows: 20.19-12.77 g/mL, and the expression between different groups is obviously different (p < 0.001). This indicates that: the APOA1 is a protein closely related to the prognosis after liver transplantation, and can be used as a primary evaluation and detection index of the effect after liver transplantation.
Therefore, the APOA1 expression of the serum sample to be detected can be preliminarily judged to be a patient (7.08-15.13ug/mL) with good postoperative recovery before liver transplantation (4.74-11.7 ug/mL) or a normal healthy population (20.19-12.77 ug/mL) through an ELISA experiment.
The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.
Sequence listing
<110> first subsidiary Hospital of medical college of Western-Ann transportation university
<120> application of diagnosis marker APOA1 for evaluating effect after liver transplantation
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>1
Ser Pro Pro Met Gly Leu Val Val Ala Pro Thr Gly Leu
1 5 10
Claims (6)
1. An application of a serum polypeptide molecular diagnostic marker APOA1 in preparing a diagnostic reagent for evaluating the effect after liver transplantation is characterized in that the amino acid sequence of the serum polypeptide molecular diagnostic marker APOA1 is shown in SEQ ID No. 1.
2. The use of claim 1, wherein the serum polypeptide molecular diagnostic marker APOA1 is apolipoprotein A1 with a molecular weight of 2997.64 daltons.
3. The use of claim 1, wherein the serum polypeptide molecular diagnostic marker APOA1 has a detection parameter in serum ranging from 8.99 ug/mL to 13.07 ug/mL.
4. The use of claim 1, wherein the diagnostic reagent for evaluation of the effect after liver transplantation is a reagent for detecting serum polypeptide molecules for evaluation of the effect after liver transplantation by ELISA.
5. Use of a molecule that binds to the serum polypeptide molecular diagnostic marker APOA1 of claim 1 in the preparation of a diagnostic reagent for evaluating the effect after liver transplantation.
6. Application of molecules combined with APOA1 protein in preparation of diagnostic reagents for evaluating effects after liver transplantation.
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Non-Patent Citations (3)
| Title |
|---|
| CLAUDE L. MALRNENDIERA等: "Lipid and apolipoprotein changes after orthotopic liver transplantation for end-stage liver diseases", 《CLINICA CHIMICA ACTA》 * |
| JOHN D. PALOMBO等: "Effectiveness of Orthotopic Liver Transplantation on the Restoration of Cholesterol Metabolism in Patients With End-Stage Liver Disease", 《GASTROENTEROLOGY》 * |
| V. W. ARMSTRONG等: "Relationship of apolipoproteins Al, B and lipoprotein Lp(a) to hepatic function of liver recipients during the early post-transplant period", 《EUROPEAN JOURNAL OF CLINICAL INVESTIGATION》 * |
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