CN113999841A - Protein scaffold OVAL100 and application thereof in radioligand method - Google Patents
Protein scaffold OVAL100 and application thereof in radioligand method Download PDFInfo
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- 238000000034 method Methods 0.000 title abstract description 22
- 239000000427 antigen Substances 0.000 claims abstract description 33
- 102000036639 antigens Human genes 0.000 claims abstract description 33
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- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 7
- 238000003556 assay Methods 0.000 claims abstract description 5
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- 239000013612 plasmid Substances 0.000 claims description 13
- 229960004452 methionine Drugs 0.000 claims description 10
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims description 8
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- 238000003653 radioligand binding assay Methods 0.000 description 11
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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Abstract
The invention discloses a protein scaffold OVAL100 capable of carrying radioactive signals and application thereof in detecting antibodies by a radioligand method, belonging to the technical field of biology. The nucleotide sequence of the protein scaffold OVAL100 is shown in SEQ ID NO. 1. The protein scaffold OVAL100 of the present invention is effective in carrying a radioactive signal in RBA assays, the radiation readings of which increase with increasing antibody content; the OVAL100 protein scaffold can be used for carrying tiny antigens and some antigens which cannot carry radioactive signals by themselves, and is used for capturing and identifying antibodies.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a protein scaffold OVAL100 capable of carrying radioactive signals and application thereof in detecting antibodies by a radioligand method.
Background
The radioligand method has the advantages of sensitive signal, wide linear range, low cost and the like in the detection of antigen and antibody. Detection of ZnT8 autoantibodies using Radioligand Binding Assay (RBA) of the receptor,has become the gold standard for detecting diabetes autoantibodies in the world at present. The method has unique advantages in the antigen preparation process: 1. replacing methionine containing S in antigen with35The S-labeled radioactive methionine does not change any space structure of the original antigen and is different from other detection methods needing to label foreign substances; 2. the preparation of the rapid transcription translation antigen only needs 90 minutes, the time is short, and the cost is low; 3. the amount of antigen used is extremely low, and thousands of samples can be detected by the antigen prepared in 90 minutes.
However, the inventors have found in their studies that radioactive antigens need to satisfy certain conditions during their preparation: 1) the sequence of the antigen needs to contain a certain amount of methionine, but part of the antigen does not contain or contains a very small amount of methionine, and cannot effectively carry radioactive signals, so that the radioactive antigen cannot be quickly transcribed and translated, and the application of the method is limited; 2) part of the minute antigenic domains, by themselves too small, are not only difficult to routinely biosynthesize, but also incapable of rapid transcriptional translation.
Disclosure of Invention
The invention aims to provide a protein scaffold OVAL100 capable of carrying a radioactive signal, which is rich in methionine, can be connected with a known antigen, obtains radioactive antigen through transcription and translation and captures an antibody to complete the detection by a radioligand method.
In order to achieve the purpose, the invention adopts the following technical scheme:
the nucleotide sequence of the protein scaffold OVAL100 is shown in SEQ ID NO. 1. The method comprises the following specific steps:
GTGACTGAGCAAGAAAGCAAACCTGTGCAGATGATGTACCAGATTGGTTTATTTAGAGTGGCATCAATGGCTTCTGAGAAAATGAAGATCCTGGAGCTTCCATTTGCCAGTGGGACAATGAGCATGTTGGTGCTGTTGCCTGATGAAGTCTCAGGCCTTGAGCAGCTTGAGAGTATAATCAACTTTGAAAAACTGACTGAATGGACCAGTTCTAATGTTATGGAAGAGAGGAAGATCAAAGTGTACTTACCTCGCATGAAGATGGAGGAAAAATACAACCTCACATCTGTCTTAATGGCT。
the protein scaffold OVAL100 is applied to detecting antibodies by a radioligand method.
Further, the antibody is a ZnT8 autoantibody.
Further, the specific steps of the application are as follows:
step 2, adopt35Marking the plasmid obtained in the step 1 by S-methionine, and performing transcription and translation to obtain a radioactive marked antigen;
and 3, detecting the antibody to be detected by adopting the antigen obtained in the step 2.
The protein scaffold OVAL100 of the present invention is effective in carrying a radioactive signal in RBA assays, the radiation readings of which increase with increasing antibody content; the OVAL100 protein scaffold can be used for carrying tiny antigens and some antigens which cannot carry radioactive signals by themselves, and is used for capturing and identifying antibodies.
Drawings
FIG. 1 shows the efficiency of a protein scaffold OVAL100 to carry a radioactive signal.
FIG. 2 shows ROC curves for RBA detection of ZnT8 and OVAL100ZnT 8.
FIG. 3 shows the correlation analysis results of RBA detection ZnT8 and OVAL100ZnT 8.
Detailed Description
The present invention is described in further detail below with reference to specific examples, but the present invention should not be construed as being limited thereto. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention. The experimental methods and reagents of the formulations not specified in the examples are in accordance with the conventional conditions in the art.
Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.
Example 1
Application of protein scaffold OVAL100 in detection of ZnT8 autoantibody by radioligand method
1. Experimental Material
(1) Sample source: ZnT8 positive control serum samples were taken from Barbara Davis Center for Childhood Diabetes laboratories, USA, and negative control serum samples were taken from healthy volunteers without family history of Diabetes. 192 blood samples from patients with diabetes are obtained from patients with clinical blood sugar abnormality. Type 1 diabetes (T1 DM) ZnT8 positive sera in 17 cases, from the clinical diagnostic T1D samples validated by the applicant's RBA ZnT8 test technology platform. 133 healthy persons (Health) were from the recruited population [ age (23.8 ± 8.9) years; 72 male and 61 female cases ]; the glucose tolerance test (OGTT) is fasting and the blood sugar is normal for 2h, chronic and endocrine diseases such as heart, brain, liver, kidney and the like are excluded, and family history of diabetes and autoimmune disease history are not existed.
(2) Main reagents and instruments: TNT SP6 rapid transcription translation kit (Promega corporation, cat # L2080);35s-methionine (5 mCi, PerkinElmer, cat # NEG 709A); protein A Sepharose (GE, cat # 17-5280-02); NAP-5 column (GE Co.); 96-well PVDF plate (Corning Corp., Cat. 3504); microscint-20 scintillation fluid (PerkinElmer, Cat. 6013621); TBST buffer (Tris-Base 2.424g, NaCl 8.70g, Tween-201.5 mL, BSA 1.0g, distilled water to 1000mL, pH 7.4); β Counter liquid scintillation Counter (PerkinElmer); protein purification Column NAP Column (GE, cat # 17-0853-02); Anti-Ovalbumin antibody (abcam, cat # ab 181688).
2. Experimental methods
(1) Construction of three plasmids
OVAL100 
ZnT8 
OVAL100ZnT8
a. The construction scheme is as follows: after codon optimization is carried out according to a mammalian protein expression system (two enzyme cutting sites are avoided), the gene is constructed on a vector and comprises a kozak sequence, XhoI and XbaI enzyme cutting sites and a target sequence. The method is as follows:
XhoI (ctcgag) + kozak sequence (gccacc) + ATG + order sequence + stop codon + XbaI
b. The target sequence is as follows:
plasmids
The sequence of OVAL100 is shown in SEQ ID NO. 2. The method comprises the following specific steps:
GTGACTGAGCAAGAAAGCAAACCTGTGCAGATGATGTACCAGATTGGTTTATTTAGAGTGGCATCAATGGCTTCTGAGAAAATGAAGATCCTGGAGCTTCCATTTGCCAGTGGGACAATGAGCATGTTGGTGCTGTTGCCTGATGAAGTCTCAGGCCTTGAGCAGCTTGAGAGTATAATCAACTTTGAAAAACTGACTGAATGGACCAGTTCTAATGTTATGGAAGAGAGGAAGATCAAAGTGTACTTACCTCGCATGAAGATGGAGGAAAAATACAACCTCACATCTGTCTTAATGGCTCCTAAACCCTCCACCCCGCCTGGTTCTTCAGGCGGTGGATCCGTGACTGAGCAAGAAAGCAAACCTGTGCAGATGATGTACCAGATTGGTTTATTTAGAGTGGCATCAATGGCTTCTGAGAAAATGAAGATCCTGGAGCTTCCATTTGCCAGTGGGACAATGAGCATGTTGGTGCTGTTGCCTGATGAAGTCTCAGGCCTTGAGCAGCTTGAGAGTATAATCAACTTTGAAAAACTGACTGAATGGACCAGTTCTAATGTTATGGAAGAGAGGAAGATCAAAGTGTACTTACCTCGCATGAAGATGGAGGAAAAATACAACCTCACATCTGTCTTAATGGCT
plasmids
The target sequence of ZnT8 is shown in SEQ ID NO. 3. The method comprises the following specific steps:
ATGAAGGACTTCTCCATCTTACTCATGGAAGGTGTGCCAAAGAGCCTGAATTACAGTGGTGTGAAAGAGCTTATTTTAGCAGTCGACGGGGTGCTGTCTGTGCACAGCCTGCACATCTGGTCTCTAACAATGAATCAAGTAATTCTCTCAGCTCATGTTGCTACAGCAGCCAGCCGGGACAGCCAAGTGGTTCGGAGAGAAATTGCTAAAGCCCTTAGCAAAAGCTTTACGATGCACTCACTCACCATTCAGATGGAATCTCCAGTTGACCAGGACCCCGACTGCCTTTTCTGTGAAGACCCCTGTGACCCTAAACCCTCCACCCCGCCTGGTTCTTCAGGCGGTGGATCCATGAAGGACTTCTCCATCTTACTCATGGAAGGTGTGCCAAAGAGCCTGAATTACAGTGGTGTGAAAGAGCTTATTTTAGCAGTCGACGGGGTGCTGTCTGTGCACAGCCTGCACATCTGGTCTCTAACAATGAATCAAGTAATTCTCTCAGCTCATGTTGCTACAGCAGCCAGCTGGGACAGCCAAGTGGTTCGGAGAGAAATTGCTAAAGCCCTTAGCAAAAGCTTTACGATGCACTCACTCACCATTCAGATGGAATCTCCAGTTGACCAGGACCCCGACTGCCTTTTCTGTGAAGACCCCTGTGAC
plasmids
The sequence of OVAL100ZnT8 is shown in SEQ ID NO. 4. The method comprises the following specific steps:
ATGAAGGACTTCTCCATCTTACTCATGGAAGGTGTGCCAAAGAGCCTGAATTACAGTGGTGTGAAAGAGCTTATTTTAGCAGTCGACGGGGTGCTGTCTGTGCACAGCCTGCACATCTGGTCTCTAACAATGAATCAAGTAATTCTCTCAGCTCATGTTGCTACAGCAGCCAGCCGGGACAGCCAAGTGGTTCGGAGAGAAATTGCTAAAGCCCTTAGCAAAAGCTTTACGATGCACTCACTCACCATTCAGATGGAATCTCCAGTTGACCAGGACCCCGACTGCCTTTTCTGTGAAGACCCCTGTGACCCTAAACCCTCCACCCCGCCTGGTTCTTCAGGCGGTGGATCCGTGACTGAGCAAGAAAGCAAACCTGTGCAGATGATGTACCAGATTGGTTTATTTAGAGTGGCATCAATGGCTTCTGAGAAAATGAAGATCCTGGAGCTTCCATTTGCCAGTGGGACAATGAGCATGTTGGTGCTGTTGCCTGATGAAGTCTCAGGCCTTGAGCAGCTTGAGAGTATAATCAACTTTGAAAAACTGACTGAATGGACCAGTTCTAATGTTATGGAAGAGAGGAAGATCAAAGTGTACTTACCTCGCATGAAGATGGAGGAAAAATACAACCTCACATCTGTCTTAATGGCTCCTAAACCCTCCACCCCGCCTGGTTCTTCAGGCGGTGGATCCATGAAGGACTTCTCCATCTTACTCATGGAAGGTGTGCCAAAGAGCCTGAATTACAGTGGTGTGAAAGAGCTTATTTTAGCAGTCGACGGGGTGCTGTCTGTGCACAGCCTGCACATCTGGTCTCTAACAATGAATCAAGTAATTCTCTCAGCTCATGTTGCTACAGCAGCCAGCTGGGACAGCCAAGTGGTTCGGAGAGAAATTGCTAAAGCCCTTAGCAAAAGCTTTACGATGCACTCACTCACCATTCAGATGGAATCTCCAGTTGACCAGGACCCCGACTGCCTTTTCTGTGAAGACCCCTGTGAC
(2) transcription of the translation plasmid to obtain a radiolabeled antigen
35The mixture of S-methionine and TNT SP6 was thawed and placed on ice, and 40. mu.L of TNT mixture, 1. mu.L (1. mu.g/. mu.L) of plasmid, and 5. mu.L of plasmid were added in that order35S-methionine is supplemented with 4. mu.L nuclease-free water until the total reaction system is 50. mu.L, mixed well, placed in a 30 ℃ water bath box for incubation for 90min, and then taken out and placed on ice to prepare NAP-5 columns.
1 NAP-5 column was taken out and placed on a test tube rack, the upper and lower lids were opened, the equilibration solution was discarded, 1mL of TBST buffer was added to equilibrate the NAP-5 column, and elution was carried out 3 times. The reaction mixture was carefully applied to the surface of the NAP-5 column packing, the reaction tube was washed with 100 μ L of buffer and then applied to the NAP-5 column, after the red liquid slowly moved down to the column 2/3, 500 μ L of buffer was added, the color change of the liquid drop under the column was carefully observed, about 500 μ L of red column-passing liquid was collected, 2 μ L of column-passing antigen was taken out therefrom and mixed with 1mL of scintillation liquid in a scintillation vial, and the number of pulses per minute (CPM) was counted on a 96-well β Counter scintillation Counter.
The three proteins OVAL100, ZnT8 and OVAL100ZnT8 are transcribed and translated according to the three plasmids.
(3) Binding and detection of a test sample to an antigen
Adding 5 mu L of serum into each hole, wherein each sample and quality control serum are double-hole, taking a proper amount of labeled antigen, diluting the labeled antigen to 20000CPM/60 mu L by using 6mL of TBST buffer solution, adding 60 mu L of diluted labeled antigen into each hole, wherein the CPM value of each hole is required to be more than or equal to 20000, and uniformly mixing the labeled antigen and the serum and oscillating for 1 hour. In a refrigerator at 4 ℃ overnight. The next day, 50 μ L of the mixed solution was sequentially taken out from each well of a 96-well plate and transferred to a 96-well PVDF filter plate, 25 μ L of protein A-agarose was added to each well to precipitate an antigen-antibody complex, the antigen-antibody complex was further mixed in a refrigerator at 4 ℃ for 1 hour and then taken out, 200 μ L of TBST buffer was added to each well of the PVDF filter plate to wash the precipitate, the liquid was pumped by a vacuum pump to retain the precipitate (the pressure of the vacuum pump should not be too high in order to prevent the precipitate from being lost during pumping), 150 μ L of buffer was added to repeat washing for 7 times, the mixture was placed in an oven to dry and 200 μ L of scintillation fluid was added to each well, the mixture was counted on a 96-well β Counter, and 1min was counted per well.
And (4) calculating a result:
index = (specimen serum CPM-negative quality control CPM)/(positive quality control CPM-negative quality control CPM).
Statistical treatment:
all data were counted using GraphPad software, all measurements were normally distributed as mean ± standard deviation (x ± s), inter-group comparisons were performed using t-test, one-way variance analysis, analysis of variance trend test, comparison of both methods was performed using Receiver Operating Characteristics (ROC) curve analysis, and using Cohen's kappa coefficient analysis for consistency of results, correlation of Spearman analysis results.P<A difference of 0.05 is statistically significant.
3. Results of the experiment
(1) Effectiveness of protein scaffold OVAL100 to carry a radioactive signal
Transcription and translation plasmid
OVAL100, protein scaffold OVAL100 with radioactive signal was obtained, and the simulated sample was captured using the purchased antibody against OVAL100 protein as a simulated sample, and the radiation readings at different simulated sample volumes were measured according to the experimental method described above.
As shown in FIG. 1, the protein scaffold OVAL100 is effective in capturing antibodies, and carries a radioactive signal that varies with the concentration of the antibody.
(2) Consistency test of results of RBA detection ZnT8 and OVAL100ZnT8, ROC curve analysis
The conventional detection of the ZnT8 antibody in the serum of a T1DM patient is carried out by utilizing the prior international IASP-certified RBA (Ribose nucleic acid) detection ZnT8 antibody technical platform of the applicant. The ZnT8 protein was then replaced with OVAL100ZnT8 protein, the assay method was unchanged and the results of the identity test are shown in the table below.
By the consistency test, Kappa =0.932, the consistency rate of positive judgment reaches 97.95%, and the detection results of ZnT8 and OVAL100ZnT8 antibodies by the radioligand method are highly consistent.
The ROC curve is shown in fig. 2, the sensitivity of OVAL100ZnT8 antibody is 90.8% (59/65), the specificity is 99.6% (276/277), the area under the AUC curve is 0.904, P is < 0.0001, and there is no significant difference between the two.
(3) Correlation analysis of results of ZnT8 and OVAL100ZnT8 detected by RBA
192 DM blood samples, 17T 1DM ZnT8 positive sera and 133 healthy persons (Health) were respectively tested by RBA ZnT8 and OVAL100ZnT8, and the test results were analyzed for correlation when n is 342 in total.
The results are shown in FIG. 3, and the correlation r of the results of the two methods is 0.5390, 95% confidence interval 0.4567-0.6122, P < 0.0001, and the results of the two groups are highly correlated by Spearman analysis.
The protein scaffold OVAL100 of the present invention is effective in carrying a radioactive signal in RBA assays, the radiation readings of which increase with increasing antibody content; the OVAL100 can carry an antigen for capture and detection of antibodies, and in the detection of the ZnT8 antibody, the results of the two are highly correlated whether or not carrying a scaffold. Thus the OVAL100 protein scaffold can be used to carry small antigens, and some antigens which cannot carry radioactive signals by themselves, for capture and identification of antibodies.
Sequence listing
<110> Jiangsu province national hospital (the first subsidiary hospital of Nanjing medical university)
<120> protein scaffold OVAL100 and its use in radioligand method
<130> 20210916
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gcatcaatgg cttctgagaa aatgaagatc ctggagcttc catttgccag tgggacaatg 120
agcatgttgg tgctgttgcc tgatgaagtc tcaggccttg agcagcttga gagtataatc 180
aactttgaaa aactgactga atggaccagt tctaatgtta tggaagagag gaagatcaaa 240
gtgtacttac ctcgcatgaa gatggaggaa aaatacaacc tcacatctgt cttaatggct 300
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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gtgactgagc aagaaagcaa acctgtgcag atgatgtacc agattggttt atttagagtg 60
gcatcaatgg cttctgagaa aatgaagatc ctggagcttc catttgccag tgggacaatg 120
agcatgttgg tgctgttgcc tgatgaagtc tcaggccttg agcagcttga gagtataatc 180
aactttgaaa aactgactga atggaccagt tctaatgtta tggaagagag gaagatcaaa 240
gtgtacttac ctcgcatgaa gatggaggaa aaatacaacc tcacatctgt cttaatggct 300
cctaaaccct ccaccccgcc tggttcttca ggcggtggat ccgtgactga gcaagaaagc 360
aaacctgtgc agatgatgta ccagattggt ttatttagag tggcatcaat ggcttctgag 420
aaaatgaaga tcctggagct tccatttgcc agtgggacaa tgagcatgtt ggtgctgttg 480
cctgatgaag tctcaggcct tgagcagctt gagagtataa tcaactttga aaaactgact 540
gaatggacca gttctaatgt tatggaagag aggaagatca aagtgtactt acctcgcatg 600
aagatggagg aaaaatacaa cctcacatct gtcttaatgg ct 642
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atgaaggact tctccatctt actcatggaa ggtgtgccaa agagcctgaa ttacagtggt 60
gtgaaagagc ttattttagc agtcgacggg gtgctgtctg tgcacagcct gcacatctgg 120
tctctaacaa tgaatcaagt aattctctca gctcatgttg ctacagcagc cagccgggac 180
agccaagtgg ttcggagaga aattgctaaa gcccttagca aaagctttac gatgcactca 240
ctcaccattc agatggaatc tccagttgac caggaccccg actgcctttt ctgtgaagac 300
ccctgtgacc ctaaaccctc caccccgcct ggttcttcag gcggtggatc catgaaggac 360
ttctccatct tactcatgga aggtgtgcca aagagcctga attacagtgg tgtgaaagag 420
cttattttag cagtcgacgg ggtgctgtct gtgcacagcc tgcacatctg gtctctaaca 480
atgaatcaag taattctctc agctcatgtt gctacagcag ccagctggga cagccaagtg 540
gttcggagag aaattgctaa agcccttagc aaaagcttta cgatgcactc actcaccatt 600
cagatggaat ctccagttga ccaggacccc gactgccttt tctgtgaaga cccctgtgac 660
<210> 4
<211> 1002
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
atgaaggact tctccatctt actcatggaa ggtgtgccaa agagcctgaa ttacagtggt 60
gtgaaagagc ttattttagc agtcgacggg gtgctgtctg tgcacagcct gcacatctgg 120
tctctaacaa tgaatcaagt aattctctca gctcatgttg ctacagcagc cagccgggac 180
agccaagtgg ttcggagaga aattgctaaa gcccttagca aaagctttac gatgcactca 240
ctcaccattc agatggaatc tccagttgac caggaccccg actgcctttt ctgtgaagac 300
ccctgtgacc ctaaaccctc caccccgcct ggttcttcag gcggtggatc cgtgactgag 360
caagaaagca aacctgtgca gatgatgtac cagattggtt tatttagagt ggcatcaatg 420
gcttctgaga aaatgaagat cctggagctt ccatttgcca gtgggacaat gagcatgttg 480
gtgctgttgc ctgatgaagt ctcaggcctt gagcagcttg agagtataat caactttgaa 540
aaactgactg aatggaccag ttctaatgtt atggaagaga ggaagatcaa agtgtactta 600
cctcgcatga agatggagga aaaatacaac ctcacatctg tcttaatggc tcctaaaccc 660
tccaccccgc ctggttcttc aggcggtgga tccatgaagg acttctccat cttactcatg 720
gaaggtgtgc caaagagcct gaattacagt ggtgtgaaag agcttatttt agcagtcgac 780
ggggtgctgt ctgtgcacag cctgcacatc tggtctctaa caatgaatca agtaattctc 840
tcagctcatg ttgctacagc agccagctgg gacagccaag tggttcggag agaaattgct 900
aaagccctta gcaaaagctt tacgatgcac tcactcacca ttcagatgga atctccagtt 960
gaccaggacc ccgactgcct tttctgtgaa gacccctgtg ac 1002
Claims (4)
1. The nucleotide sequence of the protein scaffold OVAL100 is shown in SEQ ID NO. 1.
2. Use of the protein scaffold OVAL100 of claim 1 in the detection of antibodies by radioligand assay.
3. Use according to claim 2, characterized in that: the antibody is a ZnT8 autoantibody.
4. Use according to claim 2, characterized in that: the application comprises the following specific steps:
step 1, constructing a plasmid containing a protein scaffold OVAL100 nucleotide sequence and an antigen nucleotide sequence;
step 2, adopt35Marking the plasmid obtained in the step 1 by S-methionine, and performing transcription and translation to obtain a radioactive marked antigen;
and 3, detecting the antibody to be detected by adopting the antigen obtained in the step 2.
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| CN202111085878.6A CN113999841A (en) | 2021-09-16 | 2021-09-16 | Protein scaffold OVAL100 and application thereof in radioligand method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117417427A (en) * | 2023-09-11 | 2024-01-19 | 江苏省人民医院（南京医科大学第一附属医院） | Kit for detecting endogenous autologous insulin-mediated insulin antibody |
| CN117434270A (en) * | 2023-09-11 | 2024-01-23 | 江苏省人民医院（南京医科大学第一附属医院） | Protein scaffold capable of carrying radioactive signals and application of protein scaffold in antibody detection |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101646781A (en) * | 2006-12-29 | 2010-02-10 | 科罗拉多大学董事会 | The diagnosis of autoimmune disease and treatment target and uses thereof |
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2021
- 2021-09-16 CN CN202111085878.6A patent/CN113999841A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101646781A (en) * | 2006-12-29 | 2010-02-10 | 科罗拉多大学董事会 | The diagnosis of autoimmune disease and treatment target and uses thereof |
Non-Patent Citations (1)
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| CHARLES LANE等: "Sequestration and Turnover of Guinea-pig Milk Proteins and Chicken Ovalbumin in Xenopus Oocytes", 《EUR. J. BIOCHEM.》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117417427A (en) * | 2023-09-11 | 2024-01-19 | 江苏省人民医院（南京医科大学第一附属医院） | Kit for detecting endogenous autologous insulin-mediated insulin antibody |
| CN117434270A (en) * | 2023-09-11 | 2024-01-23 | 江苏省人民医院（南京医科大学第一附属医院） | Protein scaffold capable of carrying radioactive signals and application of protein scaffold in antibody detection |
| CN117434270B (en) * | 2023-09-11 | 2024-05-07 | 江苏省人民医院（南京医科大学第一附属医院） | Protein scaffold capable of carrying radioactive signals and application of protein scaffold in antibody detection |
| CN117417427B (en) * | 2023-09-11 | 2024-05-07 | 江苏省人民医院（南京医科大学第一附属医院） | Kit for detecting endogenous autologous insulin-mediated insulin antibody |
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