CN102162813A - Reagent kit and method for detecting T-2 toxin by using genetically engineered single-chain antibody - Google Patents
Reagent kit and method for detecting T-2 toxin by using genetically engineered single-chain antibody Download PDFInfo
- Publication number
- CN102162813A CN102162813A CN2011100228137A CN201110022813A CN102162813A CN 102162813 A CN102162813 A CN 102162813A CN 2011100228137 A CN2011100228137 A CN 2011100228137A CN 201110022813 A CN201110022813 A CN 201110022813A CN 102162813 A CN102162813 A CN 102162813A
- Authority
- CN
- China
- Prior art keywords
- reagent
- toxin
- sample
- hole
- chain antibody
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides a reagent kit and a method for detecting T-2 toxin by using a genetically engineered single-chain antibody. Reagents in the reagent kit comprise a sample extract, a standard reagent, an enzyme labeled antigen reagent, cleaning solution, a bovine serum albumin (BSA) reagent, a chromogenic substrate and stop solution. The content of the T-2 toxin in a sample is calculated through sample processing, reagent balancing, washing, sample loading, washing, color developing, and stopping. The T-2 toxin single-chain antibody used in the reagent kit can be efficiently expressed in Escherichia coli, and can be produced on a large scale; and the preparation process is simple, practicable, time-saving, labor-saving and money-saving. By the reagent kit for detecting the T-2 toxin, whether the sample is polluted by the T-2 toxin can be determined within 1.5 to 2 hours, and the amount of the contained T-2 toxin can be calculated. The reagent kit is convenient and fast to use, and low in cost.
Description
Technical field
The present invention relates to a kind of detection kit of utilizing phage single chain antibody to detect mycotoxin, related more specifically to utilize phage single chain antibody to detect the kit of T-2 toxin, further relate to the method for using this kit to detect the T-2 toxin.
Background technology
At present, the antibody that is used for immune detection T-2 toxin all is monoclonal antibody.Antigen-immunized animal is adopted in the production of monoclonal antibody, utilizes splenocyte of immune animal and myeloma cell to merge the formation hybridoma then, filters out at last to have high antibody activity and the prolific hybridoma of energy.The whole process of production complexity, the time of consumption is long, and expense height, especially operating process need skilled professional and technical personnel to be competent at.
Summary of the invention
When overcoming existing monoclonal antibody manufacturing cost, effort, expensive deficiency, the invention provides kit and method that a kind of phage single chain antibody detects the T-2 toxin, make that the testing process of T-2 toxin is simple and easy to do, time saving and energy saving saving money,
Technical scheme of the present invention is as follows:
The kit that utilizes phage single chain antibody to detect the T-2 toxin of the present invention, its reagent comprises:
(1) sample extracting solution: the physiological saline that contains 50% (volume ratio) dimethyl sulfoxide (DMSO);
(2) T-2 toxin standard reagent: the sample extracting solution that contains the T-2 toxin;
(3) enzyme-labelled antigen reagent: contain the PBS solution of 5000 the T-2-HRP coupling protein of tiring, be stored in the 50%(volume ratio) in the glycerine ,-20 ℃ of preservations; Dilute with BSA reagent during use;
(4) cleansing solution: TNT solution; Formula constituent: the 0.05%(volume ratio) tween-20,20 mmol/L Tris-HCl, 150 mmol/L NaCl, pH 7.4;
(5) BSA reagent: the TNT solution that contains 5% (weight ratio) BSA;
(6) chromogenic substrate: formula constituent: 250 μ L, 20 mg/mL DAB, 40 μ L, 40 mg/mL NiCl, the 1.0 mol/L Tris-HCl (pH 6.8) of 9.75 ml ,-20 ℃ of preservations; Add 7 μ L 30%H during use
2O
2
(7) stop buffer: contain in the deionized water: 0.1 mol/L sodium sulphite, 2 mol/L sulfuric acid.
Use above-mentioned phage single chain antibody to detect the method for the kit of T-2 toxin, may further comprise the steps successively:
(1) sample preparation:
In 0.5～1.5 g sample, add 2～6 ml sample extracting solutions, after liquid nitrogen or mechanical lapping are pulverized, ultrasonic extraction, centrifugal, supernatant is the extract that contains sample;
It is evenly mixed to contain the extract of sample and enzyme-labelled antigen reagent equal-volume, A reagent; When using, described enzyme-labelled antigen reagent, adds the PBS solution of 1 μ l T-2-HRP coupling protein in the 5ml BSA reagent earlier with the dilution of BSA reagent;
It is mixed evenly with enzyme-labelled antigen reagent equal-volume respectively that T-2 toxin standard reagent is got series concentration, the B reagent of series concentration;
(2) reagent balance: with the kit balance to room temperature;
(3) aperture numbering: take out enzyme mark bar and be placed on the reaction plate, negative hole, No. 1 hole of mark, the 2-6 hole is T-2 toxin standard control hole, all the other holes are sample well; Enzyme mark bar has had the anti-T-2 toxin of immobilization single-chain antibody in every hole;
(4) washing: add 200～300 μ L cleansing solutions every hole in, cleansing solution must not overfolw hole outside, behind the placement 2min, remove cleansing solution, on thieving paper, pat dry, repeated washing is once;
(5) application of sample: No. 1 the hole adds 50 μ L BSA reagent, and the 2-6 hole adds the B reagent 50 μ L of series concentration respectively, and sample well adds corresponding A reagent 50 μ L, and light shaking makes each hole mixing;
(6) reaction: reaction plate is put into 37 ℃ of constant temperature ovens hatch 30 min;
(7) washing: take out reaction plate, remove reactant liquor, pat dry; Every hole adds 200～300 μ L cleansing solutions, and cleansing solution must not overflow, place 2 min after, remove cleansing solution, on thieving paper, pat dry repeated washing 4 times;
(8) colour developing: every hole adds chromogenic substrate 100 μ L, shakes up, and reaction plate is put into 37 ℃ of constant temperature ovens deposit 15 min;
(9) termination and Instrument measuring: every hole adds stop buffer 50 μ L respectively, measure the light absorption value A in each hole then at 450 nm places with microplate reader, series concentration with B reagent is a horizontal ordinate, with its corresponding light absorption value A is ordinate drawing standard curve, obtain the concentration of T-2 toxin in the sample according to typical curve, according to computing formula T-2 content of toxins (μ g/g)=C*V/m, the concentration of C-T-2 toxin wherein, μ g/mL; The volume of V-sample extracting solution, mL; The m-sample quality, g; Thereby the content of T-2 toxin in the calculation sample.
The amino acid sequence of described anti-T-2 toxin single-chain antibody is shown in SEQ ID No.1.The preparation method of described T-2 toxin single-chain antibody is as follows: extract total RNA from the spleen of immune mouse, become cDNA through reverse transcription, through the variable region of heavy chain V of pcr amplification cDNA
HGene and variable region of light chain V
LGene passes through one section connection peptides again with V
HAnd V
LConnect, form single-chain antibody, it is cloned into makes up phage antibody library on the carrier then, again by biological elutriation with the anti-T-2 toxin single-chain antibody of acquisition to T-2 toxin tool high-affinity.The amino acid sequence of described anti-T-2 toxin single-chain antibody is shown in SEQ ID No.1.
Anti-T-2 toxin single-chain antibody of the present invention, its structure as shown in Figure 1.This single-chain antibody (scFv) be with gene engineering method with mouse cDNA variable region of heavy chain (V
H) and variable region of light chain (V
L) recombinant protein that is formed by connecting by one section connection peptides (Linker, Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser).As shown in Figure 2, variable region of heavy chain (V
H) size be 340 bp; As shown in Figure 3, variable region of light chain (V
L) size be 325 bp; As shown in Figure 4, the size of anti-T-2 toxin single-chain antibody (scFv) is 750 bp; This single-chain antibody (scFv) has kept the affine activity of antigen and the functional antibody fragment of specific minimum of parental antibody, can economy of large scale production in bacterium, make that the antibody producing that is used for immune detection is easy and economical, significantly reduced the expense of diagnostic reagent.
Remarkable advantage of the present invention:
The T-2 toxin single-chain antibody that adopts in the kit of the present invention can efficiently express in Escherichia coli, and can be mass-produced, and preparation process is simple and easy to do, time saving and energy saving saving money.Utilize kit of the present invention to detect the T-2 toxin, can determine in 1.5-2 h whether sample is subjected to the T-2 endotoxin contamination, and the amount of calculating contained T-2 toxin, convenient to use, with low cost.
Description of drawings
Fig. 1 is the structural representation of anti-T-2 toxin single-chain antibody of the present invention.
Fig. 2 is heavy chain gene V
HThe electrophoretogram of amplification.
Fig. 3 is light chain gene V
LThe electrophoretogram of amplification.
Fig. 4 is the electrophoretogram of anti-T-2 toxin single-chain antibody gene scFv amplification of the present invention.
Fig. 5 is anti-T-2 toxin single-chain antibody competitive ELISA detection curve figure of the present invention.
Embodiment
Be instantiation of the present invention below, further describe the present invention, but the present invention be not limited only to this.
Make the kit that utilizes phage single chain antibody to detect the T-2 toxin by following prescription:
(1) sample extracting solution: the physiological saline of 50% volume ratio DMSO;
(2) T-2 toxin standard reagent: be respectively and contain the sample extracting solution that concentration is 0.04,0.08,0.16,0.32,0.64 μ g/mL T-2 toxin;
(3) in glycerine enzyme-labelled antigen reagent: contain the 0.01M PBS solution of tiring, be stored in the 50%(volume ratio) to the 5000T-2-HRP coupling protein ,-20 ℃ of preservations; With the dilution of BSA reagent, add the PBS solution of 1 μ l T-2-HRP coupling protein in the 5ml BSA reagent during use;
(4) cleansing solution: TNT solution; Formula constituent: the 0.05%(volume ratio) Tween-20,20 mmol/L Tris-HCl, 150 mmol/L NaCl, pH 7.4;
(5) BSA reagent: the TNT solution that contains 5% BSA;
(6) chromogenic substrate: the 10 ml liquid that develops the color; Formula constituent: 250 μ L, 20 mg/mL DAB, 40 μ L, 40 mg/mL NiCl, the 1.0 mol/L Tris-HCl (pH 6.8) of 9.75 ml ,-20 ℃ of preservations; Add 7 μ L 30%H during use
2O
2
(7) stop buffer: contain in the deionized water: 0.1 mol/L sodium sulphite, 2 mol/L sulfuric acid.
Utilize the mentioned reagent box to detect the method for T-2 toxin, concrete steps are as follows:
(1) sample preparation: add the 4ml sample extracting solution in the 1g sample, liquid nitrogen or mechanical lapping are pulverized, ultrasonic extraction 10 min, and centrifugal 10 min of 5000 rpm, supernatant is the extract that contains sample;
The sample extracting solution that contains sample gets 100 μ l and 100 μ l enzyme-labelled antigen reagent are evenly mixed, A reagent; , dilution earlier before enzyme-labelled antigen reagent uses, the PBS solution of adding 1 μ l T-2-HRP coupling protein in every 5ml BSA reagent;
Each 100 μ L of T-2 toxin standard reagent 0.04,0.08,0.16,0.32, the 0.64 μ g/mL of series concentration are evenly mixed with 100 μ L enzyme-labelled antigen reagent respectively, the B reagent of series concentration;
(2) reagent balance: kit is taken out from-20 ℃ of refrigerators, place more than 15 min, balance is to room temperature;
(3) aperture numbering: take out enzyme mark bar as required and be placed on the reaction plate support.Negative hole, No. 1 hole, 2-6 hole are T-2 toxin standard control hole, and all the other holes are sample well; Enzyme mark bar has had the anti-T-2 toxin of immobilization single-chain antibody in every hole; Described anti-T-2 toxin single-chain antibody preparation process is: extract total RNA from the spleen of immune mouse, become cDNA through reverse transcription, through pcr amplification antibody heavy chain variable region V
HGene and variable region of light chain V
LGene, by one section connection peptides (Linker) V
HAnd V
LConnect into single-chain antibody (scFv), be cloned into then and make up phage antibody library on the carrier, again by biological elutriation to obtain single-chain antibody to T-2 toxin tool high-affinity.
(4) washing: every hole adds 250 μ L cleansing solutions, and washing lotion must not be overflowed, and behind the placement 2min, removes cleansing solution, pats dry on thieving paper, repeats to wash plate once;
(5) application of sample: No. 1 the hole adds 50 μ L BSA reagent, and the 2-6 hole adds the B reagent 50 μ L of series concentration respectively, and sample well adds corresponding A reagent 50 μ L, and light shaking makes each hole mixing;
(6) reaction: reaction plate is put into 37 ℃ of constant temperature ovens hatch 30 min;
(7) washing: take out reaction plate, remove reactant liquor, pat dry.Every hole adds 250 μ L cleansing solutions, and washing lotion must not be overflowed, place 2 min after, remove cleansing solution, on thieving paper, pat dry, repeat to wash plate 4 times;
(8) colour developing: every hole adds chromogenic substrate 100 μ L, shakes up, and reaction plate is put into 37 ℃ of constant temperature ovens deposit 15 min;
(9) termination and Instrument measuring: every hole adds stop buffer 50 μ L respectively, measure the light absorption value A in each hole then at 450 nm places with microplate reader, series concentration with B reagent is a horizontal ordinate, with its corresponding light absorption value A is ordinate drawing standard curve, obtain the concentration of T-2 toxin in the sample according to typical curve, according to computing formula T-2 content of toxins (μ g/g)=C*V/m, the concentration of C-T-2 toxin wherein, μ g/mL; The volume of V-sample extracting solution, mL; The m-sample quality, g; The content of T-2 toxin in the calculation sample.
With T-2 toxin single-chain antibody and the anti-E-tag antibodies that is fixed on the ELISA Plate, form insolubilized antibody by the E-tag on the T-2 toxin single-chain antibody; Add testing sample and anti-T-2 toxin enzyme-labelled antigen reagent, antigen in the sample and enzyme-labelled antigen competition combine with insolubilized antibody, antigenic content is high more in the testing sample, what then combine with insolubilized antibody is many more, the chance that makes enzyme-labelled antigen combine with insolubilized antibody is just few more, even the combination of having no chance, adding like this and just do not develop the color or develop the color behind the substrate very shallow, the dark person that develops the color is negative.As shown in table 1, along with the progressively rising of the concentration of the normal concentration T-2 toxin that is added, corresponding light absorption value OD value gradually reduces.In view of the above the T-2 toxin normal concentration curve drawn of data as shown in Figure 5, the concentration of OD450 nm place light absorption value and testing sample is inverse relation, so just can judge the concentration of the T-2 toxin that wherein contains according to the OD450 nm place light absorption value of testing sample.
As shown in table 1, analyze.According to T-2 content of toxins in the computing formula sample (μ g/g)=C*V/m, the concentration of T-2 toxin in the C-sample wherein, μ g/mL; The volume of V-sample extracting solution, mL; The m-sample quality, g; The content of T-2 toxin in the calculation sample.When the volume of sample extracting solution is 4ml, sample quality is 1g, and light absorption value is 0.946 o'clock, and the T-2 content of toxins is: 0.04*4/1=0.16 μ g/g.
The competitive ELISA of table 1 T-2 toxin of the present invention single-chain antibody detects
Annotate: when concentration is 0 μ g/mL, an anti-mixing for single-chain antibody of the present invention and isopyknic PBS, other sample wells add isopyknic T-2 toxin standard solution (concentration is respectively 0.04,0.08,0.16,0.32,0.64 μ g/mL)
＜110〉University Of Agriculture and Forestry In Fujian
＜120〉a kind of phage single chain antibody detects the kit and the method for T-2 toxin
<160>?1
<210>?1
<211>?247
<212>?PRT
＜213〉the BABA/c mouse (
Mus musculus)
<400>1
Gln?Val?Gln?Leu?Gln?Gln?Ser?Gly?Thr?Val?Leu?Ala?Arg?Pro?Gly?Thr
1 5 10 15
Ser?Val?Lys?Met?Ser?Cys?Lys?Thr?Ser?Ser?Phe?Tyr?Thr?Phe?Gly?Ala
20 25 30
Cys?Ser?Asn?Trp?Ile?Lys?Gln?Arg?Pro?Gly?Gln?Gly?Leu?Glu?Trp?Ile
35 40 45
Gly?Trp?Val?Asn?Asp?Ile?Ser?Thr?Thr?Ser?Trp?Gly?Asp?Pro?His?Pro
50 55 60
Lys?Val?Lys?Ala?Lys?Leu?Thr?Ala?Val?Thr?Ser?Thr?Asn?Thr?Ala?Tyr
65 70 75 80
Met?Glu?Val?Ser?Ser?Leu?Thr?Asn?Glu?Asp?Ser?Ala?Val?Tyr?Tyr?Cys
85 90 95
Thr?Arg?Thr?Gly?Tyr?Ser?Thr?Pro?Arg?Trp?Gln?Gly?Trp?Gly?Gln?Gly
100 105 110
Thr?Thr?Val?Thr?Val?Ser?Ser?Gly?Gly?Gly?Gly?Ser?Gly?Gly?Gly?Gly
115 120 125
Ser?Gly?Gly?Gly?Gly?Ser?Gln?Ala?Val?Val?Thr?Gln?Glu?Ser?Ala?Leu
130 135 140
Thr?Thr?Ser?Pro?Gly?Glu?Thr?Val?Thr?Leu?Thr?Cys?Pro?Thr?Thr?Ala
145 150 155 160
Asp?Thr?Ser?Tyr?Thr?Asn?Ser?Asn?Arg?Trp?Trp?Val?Gln?Glu?Lys?Pro
165 170 175
Asp?His?Leu?Phe?Thr?Ala?Leu?Ile?Gly?Thr?Asp?Asp?Gly?Arg?Phe?Ala
180 185 190
Gly?Val?Pro?Ala?Arg?Phe?Ser?Gly?Ser?Leu?Ile?Gly?Asp?Lys?Ala?Ala
195 200 205
Leu?Thr?Ile?Thr?Gly?Ala?Gln?Thr?Glu?Asp?Glu?Ala?Ile?Tyr?Phe?Phe
210 215 220
Thr?Ser?Val?Ser?Cys?Trp?Tyr?Cys?Asn?Val?Phe?Gly?Gly?Gly?Thr?Lys
225 230 235 240
Leu?Thr?Val?Leu?Gly?Gln?Pro
245
Claims (3)
1. a phage single chain antibody detects the kit of T-2 toxin, and it is characterized in that: its reagent comprises:
(1) sample extracting solution: the physiological saline that contains 50% (volume ratio) dimethyl sulfoxide (DMSO);
(2) T-2 toxin standard reagent: the sample extracting solution that contains the T-2 toxin;
(3) enzyme-labelled antigen reagent: contain the PBS solution of 5000 the T-2-HRP coupling protein of tiring, be stored in the 50%(volume ratio) in the glycerine ,-20 ℃ of preservations; Dilute with BSA reagent during use;
(4) cleansing solution: TNT solution; Formula constituent: the 0.05%(volume ratio) tween-20,20 mmol/L Tris-HCl, 150 mmol/L NaCl, pH 7.4;
(5) BSA reagent: the TNT solution that contains 5% (weight ratio) BSA;
(6) chromogenic substrate: formula constituent: 250 μ L, 20 mg/mL DAB, 40 μ L, 40 mg/mL NiCl, the 1.0 mol/L Tris-HCl (pH 6.8) of 9.75 ml ,-20 ℃ of preservations; Add 7 μ L 30%H during use
2O
2
(7) stop buffer: contain in the deionized water: 0.1 mol/L sodium sulphite, 2 mol/L sulfuric acid.
2. method of using the described phage single chain antibody of claim 1 to detect the kit of T-2 toxin is characterized in that: may further comprise the steps successively:
(1) sample preparation:
In 0.5～1.5 g sample, add 2～6 ml sample extracting solutions, after liquid nitrogen or mechanical lapping are pulverized, ultrasonic extraction, centrifugal, supernatant is the extract that contains sample;
It is evenly mixed to contain the extract of sample and enzyme-labelled antigen reagent equal-volume, A reagent; When using, described enzyme-labelled antigen reagent, adds the PBS solution of 1 μ l T-2-HRP coupling protein in the 5ml BSA reagent earlier with the dilution of BSA reagent;
It is mixed evenly with enzyme-labelled antigen reagent equal-volume respectively that T-2 toxin standard reagent is got series concentration, the B reagent of series concentration;
(2) reagent balance: with the kit balance to room temperature;
(3) aperture numbering: take out enzyme mark bar and be placed on the reaction plate, negative hole, No. 1 hole of mark, the 2-6 hole is T-2 toxin standard control hole, all the other holes are sample well; Enzyme mark bar has had the anti-T-2 toxin of immobilization single-chain antibody in every hole;
(4) washing: add 200～300 μ L cleansing solutions every hole in, cleansing solution must not overfolw hole outside, behind the placement 2min, remove cleansing solution, on thieving paper, pat dry, repeated washing is once;
(5) application of sample: No. 1 the hole adds 50 μ L BSA reagent, and the 2-6 hole adds the B reagent 50 μ L of series concentration respectively, and sample well adds corresponding A reagent 50 μ L, and light shaking makes each hole mixing;
(6) reaction: reaction plate is put into 37 ℃ of constant temperature ovens hatch 30 min;
(7) washing: take out reaction plate, remove reactant liquor, pat dry; Every hole adds 200～300 μ L cleansing solutions, and cleansing solution must not overflow, place 2 min after, remove cleansing solution, on thieving paper, pat dry repeated washing 4 times;
(8) colour developing: every hole adds chromogenic substrate 100 μ L, shakes up, and reaction plate is put into 37 ℃ of constant temperature ovens deposit 15 min;
(9) termination and Instrument measuring: every hole adds stop buffer 50 μ L respectively, measure the light absorption value A in each hole then at 450 nm places with microplate reader, series concentration with B reagent is a horizontal ordinate, with its corresponding light absorption value A is ordinate drawing standard curve, obtain the concentration of T-2 toxin in the sample according to typical curve, according to computing formula T-2 content of toxins (μ g/g)=C*V/m, the concentration of C-T-2 toxin wherein, μ g/mL; The volume of V-sample extracting solution, mL; The m-sample quality, g; Thereby the content of T-2 toxin in the calculation sample.
3. phage single chain antibody according to claim 2 detects the using method of the kit of T-2 toxin, and it is characterized in that: the amino acid sequence of described anti-T-2 toxin single-chain antibody is shown in SEQ ID No.1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110022813.7A CN102162813B (en) | 2011-01-20 | 2011-01-20 | Reagent kit and method for detecting T-2 toxin by using genetically engineered single-chain antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110022813.7A CN102162813B (en) | 2011-01-20 | 2011-01-20 | Reagent kit and method for detecting T-2 toxin by using genetically engineered single-chain antibody |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102162813A true CN102162813A (en) | 2011-08-24 |
| CN102162813B CN102162813B (en) | 2014-12-10 |
Family
ID=44464148
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110022813.7A Expired - Fee Related CN102162813B (en) | 2011-01-20 | 2011-01-20 | Reagent kit and method for detecting T-2 toxin by using genetically engineered single-chain antibody |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102162813B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105021593A (en) * | 2015-06-12 | 2015-11-04 | 青岛科技大学 | Method for determining T-2 toxin based on foot point domain and hybridization chain reaction |
| CN105548119A (en) * | 2016-01-24 | 2016-05-04 | 湖南科技大学 | Method for rapidly detecting T-2 toxin |
| CN112798675A (en) * | 2019-11-13 | 2021-05-14 | 株式会社岛津制作所 | Method for detecting verotoxin |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101231291A (en) * | 2007-01-25 | 2008-07-30 | 天津科技大学 | Kit and method for quantitative determination of Fumonisin B1 content in food by ELISA detection technique |
| CN101699293A (en) * | 2009-10-28 | 2010-04-28 | 江苏省原子医学研究所 | Time-resolved fluoroimmunoassay kit for detecting fumonisins B1 and detection method thereof |
| CN101881771A (en) * | 2010-05-15 | 2010-11-10 | 福建农林大学 | Kit and method for detecting aflatoxin (AF)B1 of gene engineering single chain antibody |
| US20100330235A1 (en) * | 2009-06-29 | 2010-12-30 | Adiveter, S.L. | Mycotoxin adsorbent |
-
2011
- 2011-01-20 CN CN201110022813.7A patent/CN102162813B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101231291A (en) * | 2007-01-25 | 2008-07-30 | 天津科技大学 | Kit and method for quantitative determination of Fumonisin B1 content in food by ELISA detection technique |
| US20100330235A1 (en) * | 2009-06-29 | 2010-12-30 | Adiveter, S.L. | Mycotoxin adsorbent |
| CN101699293A (en) * | 2009-10-28 | 2010-04-28 | 江苏省原子医学研究所 | Time-resolved fluoroimmunoassay kit for detecting fumonisins B1 and detection method thereof |
| CN101881771A (en) * | 2010-05-15 | 2010-11-10 | 福建农林大学 | Kit and method for detecting aflatoxin (AF)B1 of gene engineering single chain antibody |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105021593A (en) * | 2015-06-12 | 2015-11-04 | 青岛科技大学 | Method for determining T-2 toxin based on foot point domain and hybridization chain reaction |
| CN105021593B (en) * | 2015-06-12 | 2017-11-28 | 青岛科技大学 | A kind of method based on foot point domain and the hybridization chain reaction measure toxin of T 2 |
| CN105548119A (en) * | 2016-01-24 | 2016-05-04 | 湖南科技大学 | Method for rapidly detecting T-2 toxin |
| CN112798675A (en) * | 2019-11-13 | 2021-05-14 | 株式会社岛津制作所 | Method for detecting verotoxin |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102162813B (en) | 2014-12-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101881771B (en) | Kit and method for detecting aflatoxin (AF)B1 of gene engineering single chain antibody | |
| Shu et al. | Anti-idiotypic nanobody: A strategy for development of sensitive and green immunoassay for Fumonisin B1 | |
| CN101993855B (en) | Hybridoma cell line 1C11 and anti-aflatoxin general monoclonal antibody generated by same as well as applications thereof | |
| CN102747042A (en) | Hybridoma cell line 10G4 and anti-alfatoxin total (B1, B2, G1 and G2) monoclonal antibody produced by the same | |
| EP2458378B1 (en) | Insulin measurement method | |
| CN102482346B (en) | Antibody specific to methicillin resistant staphylococcus aureus, detection method and kit for methicillin resistant staphylococcus aureus using the same | |
| CN109709338A (en) | Detect ox or the enzyme linked immunological kit of ovine skeletal muscle Troponin I and the preparation method and application thereof | |
| CN110526968B (en) | Staphylococcus aureus enterotoxin B nano antibody B7, application and kit | |
| CN110526966B (en) | Staphylococcus aureus enterotoxin B nano antibody B6, application and kit | |
| CN110577594B (en) | Staphylococcus aureus enterotoxin A nano antibody A21, application and kit | |
| CN111077320B (en) | ELISA kit for detecting chicken or duck skeletal muscle troponin I and preparation method and application thereof | |
| CN110563839A (en) | Staphylococcus aureus enterotoxin B nano antibody B1, application and kit | |
| CN102719405A (en) | Hybridoma cell strain 1C8 and anti-aflatoxin G1 monoclonal antibody produced by same | |
| CN102162813B (en) | Reagent kit and method for detecting T-2 toxin by using genetically engineered single-chain antibody | |
| CN102095850B (en) | Reagent kit for detecting Fumonisin B1 of genetically engineered single-chain antibody and method thereof | |
| CN108341870A (en) | A kind of anti-BSA nano antibodies, its production method and application | |
| Ji et al. | Development of a double nanobody-based sandwich immunoassay for the detecting staphylococcal enterotoxin c in dairy products | |
| CN102268091B (en) | Single-chain antibody C1B3 capable of identifying IgM (Immunoglobulin M) in grass carp | |
| CN102175847B (en) | Use method of kit for detecting tetrodotoxin with gene engineering single-chain antibody | |
| CN104597256B (en) | Article one, the peptide sequence and the application thereof that combine with bovine viral diarrhea virus E2 albumen | |
| CN103255109B (en) | Hybridoma cell strain AFG1G6, its monoclonal antibody and aflatoxin G1 immunochromatographic test strip | |
| CN102095849B (en) | Kit and method for detecting mu-conotoxin (CTX) by using single-chain antibody of gene engineering | |
| CN103328509A (en) | Method for measuring human insulin and measurement reagent | |
| CN108359642B (en) | Hybridoma cell of shrimp tropomyosin monoclonal antibody and application thereof | |
| CN103088033B (en) | The antibody gene of anti-pyrethroid pesticide scFv and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141210 Termination date: 20180120 |