CN113917151A - ELISA kit for detecting human integrin beta 4IgG antibody and application thereof - Google Patents

ELISA kit for detecting human integrin beta 4IgG antibody and application thereof Download PDF

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CN113917151A
CN113917151A CN202111051164.3A CN202111051164A CN113917151A CN 113917151 A CN113917151 A CN 113917151A CN 202111051164 A CN202111051164 A CN 202111051164A CN 113917151 A CN113917151 A CN 113917151A
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integrin beta
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CN113917151B (en
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李小光
钱华
周志军
孙小明
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract

The invention discloses an ELISA kit for detecting human integrin beta 4IgG antibody and application thereof. The kit contains integrin beta 4 microporous strips, and the integrin beta 4 microporous strips are respectively coated with polypeptides shown in SEQ ID NO. 1-39. Preferably, the kit also contains standard serum 1 and 2, enzyme labeled antibody, diluent, reaction buffer solution, washing buffer solution, enzyme substrate solution and stop solution. According to the invention, a plurality of antigen polypeptides are used for replacing a long-fragment complete protein as the coating protein of ELISA, so that the antigen epitope recognized by the antibody is fully ensured, a large amount of cost is saved, and compared with a recombinant protein, the sensitivity is higher. The invention utilizes an ELISA method to detect human integrin beta 4IgG antibody, and is more easily applied to clinical diagnosis of mucosal pemphigoid compared with the existing immunoblotting method.

Description

ELISA kit for detecting human integrin beta 4IgG antibody and application thereof
Technical Field
The invention relates to an ELISA kit and application thereof, in particular to an ELISA kit for detecting human integrin beta 4IgG antibody and application thereof. The invention belongs to the technical field of medicines.
Background
Human mucosal pemphigoid (mucous membrane pemphigoid) is a type of autoimmune blistering disease that affects mainly the skin of the elderly, mainly manifested by lesions of the oral mucosa, ocular mucosa and other mucous membranes and skin. Diagnosis cannot be made simply by clinical manifestations, and diagnosis must be confirmed by detection of autoantibodies in the serum of the patient. The major autoantigens of mucosal pemphigoid include BP180, laminin-332 and integrin α 6 β 4. Therefore, detection of autoantibodies against these autoantigens is a necessary condition for diagnosis of mucosal pemphigoid.
At present, commercial ELISA kits or reported ELISA methods can be applied to the detection of the autoantibodies against BP180 and laminin-332 in serum. However, no ELISA method for integrin beta 4 autoantibodies in serum has been reported. The existing method for detecting IgG autoantibodies of human integrin beta 4is mainly an immunoblotting method, and uses different antigen resources, including HD-rich fraction, integrin beta 4 intracellular region recombinant protein and integrin alpha 6 beta 4 extracellular region recombinant protein. Wherein the HD-rich fraction is a mixture of cell basement membrane proteins obtained after a long-term culture using a skin keratinocyte cell line (DJM-1 or A431) (1.Li X, Qian H, Sogame R, Hirako Y, Tsurta D, Ishii N, Koga H, Tsuchisaka A, Jin Z, Tsuubota K, Fukumoto A, Sotozono C, Kinoshita S, Hashimoto T. Integrin. beta.4 is amajor target antigen in Eur J tissue.2016; 26(3):247-53.(IF ═ 2.4)). However, the immunoblotting method is complicated in operation and is not suitable for clinical detection, thereby limiting its application. The acquisition of the HD-rich fraction resource is time-consuming (18 days), and the culture is carried out by using cell culture solution with relatively high price, so the cost is high; the cost of HD-rich fraction, recombinant protein in the intracellular region of integrin beta 4 and recombinant protein in the extracellular region of integrin alpha 6 beta 4 are also high.
Therefore, the invention provides an ELISA kit of integrin beta 4IgG antibody, which can detect integrin beta 4IgG autoantibody in serum of a mucosa pemphigoid patient, thereby further solving the diagnosis problem of mucosa pemphigoid.
Disclosure of Invention
The invention aims to provide an ELISA kit for detecting human integrin beta 4IgG antibody and application thereof.
In order to achieve the purpose, the invention adopts the following technical means:
the ELISA kit for detecting the human integrin beta 4IgG antibody comprises an integrin beta 4 micropore strip, wherein the integrin beta 4 micropore strip is respectively coated with polypeptides shown in SEQ ID NO. 1-39.
Preferably, the kit further comprises standard serum 1 and 2, an enzyme labeled antibody, a diluent, a reaction buffer solution, a washing buffer solution, an enzyme matrix solution and a stop solution.
Wherein, preferably, the standard serum 1 is a reaction buffer solution containing 0.05% of sodium azide w/v; the standard serum 2 is a reaction buffer solution containing 100U/ml integrin beta 4IgG antibody and 0.05% w/v sodium azide; the enzyme-labeled antibody is a rabbit anti-human IgG antibody labeled by horseradish peroxidase, the reaction buffer solution is PBS containing 0.05% v/v Tween20 and 0.05% w/v NaN3, and the cleaning buffer solution is PBS containing 0.5% v/v Tween 20; the enzyme substrate liquid is TMB color development liquid; the stop solution is 0.5N hydrochloric acid solution.
Preferably, the integrin beta 4-containing microporous band is prepared by the following method: respectively dissolving the polypeptides shown in SEQ ID NO.1-39 by PBS, storing the concentration of the polypeptides at 4mg/ml, mixing all the dissolved polypeptides with equal volume to obtain a polypeptide mixture of 4mg/ml, and diluting the polypeptide mixture by 1000 times by using PBS before coating so as to ensure that the final concentration of the polypeptide mixture reaches 4 mu g/ml; the 96-well plate was coated with the antigen polypeptide, and 100. mu.l of the polypeptide mixture was added to each well, and the coating was performed overnight at 4 ℃.
The invention relates to an IgG antibody detection kit (ELISA) for human integrin beta 4, which utilizes an ELISA method to qualitatively determine the IgG antibody of the integrin beta 4 in serum. During detection, patient serum and standard serum are added into a microporous band coated with integrin beta 4 antigen polypeptide, and the IgG antibody of integrin beta 4is combined with antigen. After washing, removing unbound serum protein, then adding an anti-human IgG antibody marked by horseradish peroxidase into the micropores to be bound with human IgG, after washing, adding a horseradish peroxidase substrate to react with the horseradish peroxidase, and finally adding an acid solution to stop the enzyme reaction. And (5) measuring the absorbance by using a microplate reader, and quantifying the detection result.
Preferably, the method for detecting the human integrin beta 4IgG antibody comprises the following steps:
(1) reagent preparation
Before the experiment, all detection materials are placed at room temperature, and a proper amount of washing buffer solution is diluted by distilled water as required;
(2) sample preparation
1:50 dilution of each patient serum: adding 500 mul reaction buffer solution into 10 mul serum, and mixing evenly;
(3) detection step
1) Adding 100 mul of diluted sample into the microporous strip, and incubating for 60 minutes at room temperature;
2) washing the microporous strip: washing 4 times with 200. mu.l/well diluted washing buffer;
3) adding 100 mul/hole enzyme-labeled antibody liquid, and incubating for 45 minutes at room temperature;
4) washing the microporous strip: washing 4 times with 200. mu.l/well diluted washing buffer;
5) adding 100 mul/hole enzyme substrate liquid, and incubating for 30 minutes at room temperature;
6) adding 100 mul/hole stop solution;
7) reading absorbance (450nm)
8) Result calculation and decision
Unit value (U/ml) calculation formula:
(A450<sample (I)>-A450<Standard serum 1>)*100/(A450<Standard serum 2>-A450<Standard serum 1>)
And (3) judging standard: unit value <14 is negative; positive results were obtained with a unit value of 14 or more.
Quality control: each test result must satisfy the following condition, otherwise the result is invalid:
standard serum 1 had an OD450 of 0.100 or less
The OD450 of the standard serum 2 was 0.500 or more.
Furthermore, the invention also provides application of the ELISA kit in preparation of a reagent for detecting the integrin beta 4IgG antibody.
Compared with the prior art, the invention has the beneficial effects that:
1) compared with the existing immunoblotting method, the ELISA method is more easily applied to clinical diagnosis of mucosal pemphigoid;
2) the integrin beta 4 antigen polypeptide is coated in the micropore strip of the kit, compared with recombinant protein, the cost is low, and the sensitivity is higher because the number of coated antigen epitopes is relatively more;
3) the invention uses a plurality of antigen polypeptides to replace long-fragment complete protein as the coating protein of ELISA, thus saving a great deal of cost while fully ensuring the antigen epitope recognized by the antibody.
Detailed Description
The present invention is further illustrated by the following experiments in conjunction with examples, it being understood that these examples are for illustrative purposes only and in no way limit the scope of the present invention.
Example 1 preparation of ELISA kit for detecting integrin beta 4IgG antibody
1. Consists of the following components:
the ELISA kit comprises:
(1) integrin beta 4 microporous strip (48 holes)
(2)0U/ml of Standard serum 1 (reaction buffer containing 0.05% sodium azide w/v, 1.5ml)
(3)100U/ml of Standard serum 2 (1.5 ml of reaction buffer containing 0.05% w/v sodium azide in which the serum of an integrin beta 4IgG autoantibody positive patient was diluted)
(4) Enzyme-labeled antibody (horse radish peroxidase-labeled rabbit anti-human IgG antibody, 8ml)
(5) Reaction buffer (0.01M PBS containing 0.05% v/vTween20 and 0.05% w/vNaN3, 50ml)
(6)10x washing buffer (0.1M PBS containing 0.5% v/vTween20, 100ml)
(7) Enzyme substrate liquid (instant TMB color developing liquid, 8ml)
(8) Stop solution (0.5N hydrochloric acid solution, 8ml)
2. Coating of integrin beta 4 microporous strips
(1) The coated material included 39 integrin β 4 antigen polypeptides as shown in table 1 below:
TABLE 139 integrin beta 4 antigen polypeptides
Figure BDA0003253008860000041
Figure BDA0003253008860000051
Figure BDA0003253008860000061
(2) Polypeptide solubilization and coating
Each polypeptide in Table 1 was solubilized with PBS at a stock concentration of 4 mg/ml. All solubilized polypeptides were mixed in equal volumes to obtain a 4mg/ml polypeptide mixture. A1000-fold dilution with PBS was performed before coating to achieve a final concentration of 4. mu.g/ml of the polypeptide mixture. The 96-well plate was coated with the antigen polypeptide by adding 100. mu.l of the polypeptide mixture (4. mu.g/ml) to each well and coating at 4 ℃ overnight (12 to 16 hours).
3. Detection method
(1) Reagent preparation
Before the experiment, all detection materials are placed at room temperature (20-30 ℃); an appropriate amount of the washing buffer (1:10) was diluted with distilled water as necessary.
(2) Sample preparation
1:50 dilution of each patient serum: mu.l of serum was added to 500. mu.l of reaction buffer and mixed well.
(3) Detection step
1) Adding 100 mul of diluted sample (1:50) into the microporous strip, and incubating for 60 minutes at room temperature (20-30 ℃);
2) washing the microporous strip: washing 4 times with 200 mul/well diluted washing buffer solution;
3) adding 100 mul/hole enzyme-labeled antibody liquid for 45 minutes at room temperature (20-30 ℃);
4) washing the microporous strip: washing 4 times with 200 mul/well diluted washing buffer solution;
5) adding 100 μ l/well enzyme matrix solution, and incubating at room temperature (20-30 deg.C) for 30 min
6) Add 100. mu.l/well stop solution
7) Reading absorbance (450nm)
8) Result calculation and decision
Unit value (U/ml) calculation formula:
(A450<sample (I)>-A450<Standard serum 1>)*100/(A450<Standard serum 2>-A450<Standard serum 1>)
And (3) judging standard: unit value <14 is negative; positive results were obtained with a unit value of 14 or more.
Quality control: each test result must satisfy the following condition, otherwise the result is invalid:
(1) standard serum 1 had an OD450 of 0.100 or less
(2) Standard serum 2 had an OD450 of 0.500 or more
Example 2 detection of Performance indicators of the kit
(1) Sample source
Serum from patients with various subtypes of cutaneous autoimmune blistering disease, a portion from the department of dermatology of the university of longstanding rice in japan, given by professor Takashi Hashimoto (patients collected from japan and all over the world), and another portion from the remaining patient serum after serological diagnosis of cutaneous autoimmune blistering disease provided by the department of dermatology of the three hospitals or the dermatology specialty hospitals in china. Normal human serum was derived from the remaining serum of a normal physical examination in a hospital.
The serum adopts 1: and (5) diluting by 50.
(2) Specificity and sensitivity
The above samples were tested according to the method established in example 1, with the results shown in table 2 below:
TABLE 2
Figure BDA0003253008860000071
(3) Repeatability of
For 5 samples, 6 replicates were performed, each with a CV% value of less than 15%.
(4) Detection range
The detection range of the kit is 5-150U/ml.
Sequence listing
<110> institute of Buddha science and technology
Primer group for loop-mediated isothermal amplification detection of Along mountain virus (ALON 120), kit containing primer group and application of kit
<160> 39
<170> SIPOSequenceListing 1.0
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<213> Homo sapiens
<400> 1
Cys Pro Ser Gly Ser Gln Arg Pro Ser Val Ser Asp Asp Thr Gly
1 5 10 15
<210> 2
<211> 15
<212> PRT
<213> Homo sapiens
<400> 2
Cys Ser Ser Asp Ala Glu Ala Pro His Gly Pro Pro Asp Asp Gly
1 5 10 15
<210> 3
<211> 15
<212> PRT
<213> Homo sapiens
<400> 3
Cys Gly Ala Gly Gly Lys Gly Gly Ser Leu Pro Arg Ser Ala Thr
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<210> 4
<211> 15
<212> PRT
<213> Homo sapiens
<400> 4
Leu Arg Glu Gly Glu Asp Lys Pro Cys Ser Gly Arg Gly Glu Cys
1 5 10 15
<210> 5
<211> 15
<212> PRT
<213> Homo sapiens
<400> 5
Gln Gly Leu Arg Val Arg Leu Arg Pro Gly Glu Glu Arg His Cys
1 5 10 15
<210> 6
<211> 15
<212> PRT
<213> Homo sapiens
<400> 6
Leu Gln Gly Glu Arg Ile Ser Gly Asn Leu Asp Ala Pro Glu Cys
1 5 10 15
<210> 7
<211> 15
<212> PRT
<213> Homo sapiens
<400> 7
Cys Thr Arg Ser Glu His Ser His Ser Thr Thr Leu Pro Arg Asp
1 5 10 15
<210> 8
<211> 15
<212> PRT
<213> Homo sapiens
<400> 8
Cys Asp Thr Pro Met Leu Arg Ser Gly Asn Leu Lys Gly Arg Asp
1 5 10 15
<210> 9
<211> 15
<212> PRT
<213> Homo sapiens
<400> 9
Gly Glu Gly Arg Tyr Glu Gly Gln Phe Cys Glu Tyr Asp Asn Cys
1 5 10 15
<210> 10
<211> 15
<212> PRT
<213> Homo sapiens
<400> 10
Gly Glu Gly Arg Tyr Glu Gly Gln Phe Cys Glu Tyr Asp Asn Cys
1 5 10 15
<210> 11
<211> 15
<212> PRT
<213> Homo sapiens
<400> 11
Cys Asp Gly Asp Ser Pro Glu Ser Arg Leu Thr Val Pro Gly Leu
1 5 10 15
<210> 12
<211> 15
<212> PRT
<213> Homo sapiens
<400> 12
Cys Lys Phe Arg Gln Gln Pro Asn Ala Gly Lys Lys Gln Asp His
1 5 10 15
<210> 13
<211> 15
<212> PRT
<213> Homo sapiens
<400> 13
Cys Asp Gln Asp Ala Arg Gly Met Val Glu Phe Gln Glu Gly Val
1 5 10 15
<210> 14
<211> 15
<212> PRT
<213> Homo sapiens
<400> 14
Cys Val Lys Ala Arg Asn Gly Ala Gly Trp Gly Pro Glu Arg Glu
1 5 10 15
<210> 15
<211> 15
<212> PRT
<213> Homo sapiens
<400> 15
Cys Val Ser Phe Glu Gln Pro Glu Phe Ser Val Ser Arg Gly Asp
1 5 10 15
<210> 16
<211> 14
<212> PRT
<213> Homo sapiens
<400> 16
Cys Asn Gly Arg Gly His Cys Glu Cys Gly Arg Cys His Cys
1 5 10
<210> 17
<211> 15
<212> PRT
<213> Homo sapiens
<400> 17
Cys Gly Pro Glu Arg Glu Gly Ile Ile Thr Ile Glu Ser Gln Asp
1 5 10 15
<210> 18
<211> 15
<212> PRT
<213> Homo sapiens
<400> 18
Cys Pro Asp Ile Pro Ile Val Asp Ala Gln Ser Gly Glu Asp Tyr
1 5 10 15
<210> 19
<211> 15
<212> PRT
<213> Homo sapiens
<400> 19
Gln Lys Thr Arg Thr Gly Ser Phe His Ile Arg Arg Gly Glu Cys
1 5 10 15
<210> 20
<211> 15
<212> PRT
<213> Homo sapiens
<400> 20
Cys Ser Pro Asp Ser Leu Gln Leu Ser Trp Glu Arg Pro Arg Arg
1 5 10 15
<210> 21
<211> 14
<212> PRT
<213> Homo sapiens
<400> 21
Cys Thr Arg Asp Ile Gly Trp Arg Pro Asp Ser Thr His Leu
1 5 10
<210> 22
<211> 15
<212> PRT
<213> Homo sapiens
<400> 22
Cys Val Leu Val His Lys Lys Lys Asp Cys Pro Pro Gly Ser Phe
1 5 10 15
<210> 23
<211> 15
<212> PRT
<213> Homo sapiens
<400> 23
Glu Glu Thr Gln Ile Asp Thr Thr Leu Arg Arg Ser Gln Met Cys
1 5 10 15
<210> 24
<211> 15
<212> PRT
<213> Homo sapiens
<400> 24
Cys Asp Asp Cys Thr Tyr Ser Tyr Thr Met Glu Gly Asp Gly Ala
1 5 10 15
<210> 25
<211> 15
<212> PRT
<213> Homo sapiens
<400> 25
Cys Ser Cys Arg Thr His Gln Glu Val Pro Ser Glu Pro Gly Arg
1 5 10 15
<210> 26
<211> 15
<212> PRT
<213> Homo sapiens
<400> 26
Ala Pro Val Lys Ser Cys Thr Glu Cys Val Arg Val Asp Lys Cys
1 5 10 15
<210> 27
<211> 15
<212> PRT
<213> Homo sapiens
<400> 27
Cys Asp Ser Glu Ser Glu Ala His Leu Leu Asp Ser Lys Val Pro
1 5 10 15
<210> 28
<211> 15
<212> PRT
<213> Homo sapiens
<400> 28
Gly Phe Leu Cys Asn Asp Arg Gly Arg Cys Ser Met Gly Gln Cys
1 5 10 15
<210> 29
<211> 15
<212> PRT
<213> Homo sapiens
<400> 29
Cys Pro Pro Glu Leu Ile Pro Arg Leu Ser Ala Ser Ser Gly Arg
1 5 10 15
<210> 30
<211> 15
<212> PRT
<213> Homo sapiens
<400> 30
Cys Leu Arg Val Ser Trp Gln Glu Pro Arg Cys Glu Arg Pro Leu
1 5 10 15
<210> 31
<211> 15
<212> PRT
<213> Homo sapiens
<400> 31
Val Asp Gly Thr His Val Cys Gln Leu Pro Glu Asp Gln Lys Cys
1 5 10 15
<210> 32
<211> 15
<212> PRT
<213> Homo sapiens
<400> 32
Cys Gly His Met Val Gly Phe Lys Glu Asp His Tyr Met Leu Arg
1 5 10 15
<210> 33
<211> 15
<212> PRT
<213> Homo sapiens
<400> 33
Ser Met Ser Asp Asp Leu Asp Asn Leu Lys Lys Met Gly Gln Cys
1 5 10 15
<210> 34
<211> 15
<212> PRT
<213> Homo sapiens
<400> 34
Cys Leu Gln Glu Val Asp Ser Leu Leu Arg Gly Arg Gln Val Arg
1 5 10 15
<210> 35
<211> 15
<212> PRT
<213> Homo sapiens
<400> 35
Cys Ala Ile Ile Asn Leu Ala Thr Gln Pro Lys Arg Pro Met Ser
1 5 10 15
<210> 36
<211> 15
<212> PRT
<213> Homo sapiens
<400> 36
Cys Ala Glu Pro Ala Glu Thr Asn Gly Glu Ile Thr Ala Tyr Glu
1 5 10 15
<210> 37
<211> 15
<212> PRT
<213> Homo sapiens
<400> 37
Cys Arg Ala Glu Glu Val Val Val Arg Cys Ser Phe Arg Asp Glu
1 5 10 15
<210> 38
<211> 15
<212> PRT
<213> Homo sapiens
<400> 38
Cys Arg Ile Pro Val Ile Arg Arg Val Leu Asp Gly Gly Lys Ser
1 5 10 15
<210> 39
<211> 15
<212> PRT
<213> Homo sapiens
<400> 39
Val Ile Ser Leu Thr Glu Asp Val Asp Glu Phe Arg Asn Lys Cys
1 5 10 15

Claims (6)

1. The ELISA kit for detecting the human integrin beta 4IgG antibody is characterized in that the kit contains an integrin beta 4 microporous band, and the integrin beta 4 microporous band is respectively coated with polypeptides shown in SEQ ID NO. 1-39.
2. The ELISA kit of claim 1 further comprising standard sera 1 and 2, enzyme-labeled antibody, diluent, reaction buffer, washing buffer, enzyme substrate solution, and stop solution.
3. The ELISA kit of claim 1 wherein the standard serum 1 is a reaction buffer containing 0.05% sodium azide w/v; the standard serum 2 is a reaction buffer solution containing 100U/ml integrin beta 4IgG antibody and 0.05% w/v sodium azide; the enzyme-labeled antibody is a rabbit anti-human IgG antibody labeled by horseradish peroxidase; the reaction buffer solution is PBS containing 0.05% v/v Tween20 and 0.05% w/v NaN3, and the washing buffer solution is PBS containing 0.5% v/v Tween 20; the enzyme substrate liquid is TMB color development liquid; the stop solution is 0.5N hydrochloric acid solution.
4. The ELISA kit of claim 1, wherein said integrin β 4-containing microwell band is prepared by the following method: respectively dissolving the polypeptides shown in SEQ ID NO.1-39 by PBS, storing the concentration of the polypeptides at 4mg/ml, mixing all the dissolved polypeptides with equal volume to obtain a polypeptide mixture of 4mg/ml, and diluting the polypeptide mixture by 1000 times by using PBS before coating so as to ensure that the final concentration of the polypeptide mixture reaches 4 mu g/ml; the 96-well plate was coated with the antigen polypeptide, and 100. mu.l of the polypeptide mixture was added to each well, and the coating was performed overnight at 4 ℃.
5. The ELISA kit of claim 1 wherein said detection of human integrin β 4IgG antibody is performed by the following steps:
(1) reagent preparation
Before the experiment, all detection materials are placed at room temperature, and a proper amount of washing buffer solution is diluted by distilled water as required;
(2) sample preparation
1:50 dilution of each patient serum: adding 500 mul reaction buffer solution into 10 mul serum, and mixing evenly;
(3) detection step
1) Adding 100 mul of diluted sample into the microporous strip, and incubating for 60 minutes at room temperature;
2) washing the microporous strip: washing 4 times with 200. mu.l/well diluted washing buffer;
3) adding 100 mul/hole enzyme-labeled antibody liquid, and incubating for 45 minutes at room temperature;
4) washing the microporous strip: washing 4 times with 200. mu.l/well diluted washing buffer;
5) adding 100 mul/hole enzyme substrate liquid, and incubating for 30 minutes at room temperature;
6) adding 100 mul/hole stop solution;
7) reading absorbance (450nm)
8) Result calculation and decision
Unit value (U/ml) calculation formula:
(A450<sample (I)>-A450<Standard serum 1>)*100/(A450<Standard serum 2>-A450<Standard serum 1>)
And (3) judging standard: unit value <14 is negative; positive results were obtained with a unit value of 14 or more.
Quality control: each test result must satisfy the following condition, otherwise the result is invalid:
standard serum 1 had an OD450 of 0.100 or less
The OD450 of the standard serum 2 was 0.500 or more.
6. Use of the ELISA kit of any of claims 1-5 in the preparation of a reagent for the detection of human integrin β 4IgG antibodies.
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JP2012233804A (en) * 2011-05-02 2012-11-29 Kurume Univ Measurement method of anti-laminin gamma-1 antibody, measurement reagent, and dermatological disease detection method using the reagent
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