CN103116024B - Application and method of anti-aflatoxin universal monoclonal antibody 1C11 in aflatoxin B1 fluorescence quenching - Google Patents
Application and method of anti-aflatoxin universal monoclonal antibody 1C11 in aflatoxin B1 fluorescence quenching Download PDFInfo
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Abstract
The invention relates to application and method of an anti-aflatoxin universal monoclonal antibody 1C11 in aflatoxin B1 fluorescence quenching. The application method for the anti-aflatoxin universal monoclonal antibody 1C11 in aflatoxin B1 fluorescence quenching comprises the step of adding the anti-aflatoxin universal monoclonal antibody 1C11 into a sample solution containing aflatoxin B1, so that aflatoxin B1 is subjected to fluorescence quenching. According to the anti-aflatoxin universal monoclonal antibody 1C11, aflatoxin B1 can be subjected to fluorescence quenching, and the fluorescence quenching can reach 90 percent; the application method is easy and controllable in operation; and moreover, the research means of biological analysis and detection of aflatoxin B1 can be enriched based on the characteristics that aflatoxin B1 can be subjected to fluorescence quenching by the anti-aflatoxin universal monoclonal antibody 1C11.
Description
Technical field
The invention belongs to field of biological detection, be specifically related to application and the method for aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in aflatoxin B1 fluorescent quenching.
Background technology
Aflatoxin is the similar compound of a class formation, and the aflatoxin of occurring in nature is mainly the secondary metabolite being produced by Aspergillus flavus and aspergillus parasiticus.The aflatoxin of finding at present has more than 20 to plant, wherein common with aflatoxin B1, B2, G1, G2 and M1.Aflatoxin can produce destruction to the liver organization of people and animal, can cause liver cancer even dead when serious.Aflatoxin delimited as I class carcinogenic substance by the World Health Organization (WHO), and the strongest with aflatoxin B1 toxicity, its toxicity is 68 times of arsenic.
The agricultural product that aflatoxin can occur in the torrid zone or subtropical zone are produced in process, also may introduce the pollution of aflatoxin in links such as storage, transportations simultaneously.Peanut is the agricultural product that the most easily find that there is aflatoxin, and other are all likely subject to aflatoxin contamination as corn, fig, kernel and cereal.Due to the cropping pattern of China smallholder decentralized, the links of agricultural product from farmland to dining table is all likely subject to the pollution of aflatoxin.And aflatoxin toxicity is large, harm is serious,
Aflatoxin, as a class natural pollutant, is different from chemical residual, near laws and regulations means, is difficult to achieve effective control.Therefore, detection technique seems particularly important in aflatoxin contamination monitoring.
The analysis of chemical residual labelled immune is a kind of conventional method of field of biological detection, and it is by adding fluorescence labeling material to carry out fluorescence labeling, then adds fluorescent quenching reagent to carry out analyzing and testing.Common fluorescent quenching reagent comprises halide ion, heavy metal ion, nitro compound, diazo-compounds etc.And consider that aflatoxin B1 self has stronger fluorescent characteristic, and easily the fluorescence of marker material is produced and disturb, therefore, never the immuno analytical method research based on marker material fluorescent quenching characteristic of aflatoxin B1 is reported.
Summary of the invention
Technical matters to be solved by this invention is to provide application and the method for aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in aflatoxin B1 fluorescent quenching for the deficiency of above-mentioned prior art existence.It can make aflatoxin B1 fluorescent quenching, and method is simple, reliable.
The present invention solves the problems of the technologies described above adopted technical scheme to be:
The application of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in aflatoxin B1 fluorescent quenching, wherein: the hybridoma cell strain 1C11 secretion that described aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 is CCTCC NO. C201013 by preserving number produces.This hybridoma cell strain has been preserved in Chinese Typical Representative culture collection center (CCTCC) on July 13rd, 2010, preservation address is, China, and Wuhan, Wuhan University, deposit number is CCTCC NO. C201013.
The application process of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in aflatoxin B1 fluorescent quenching, it is characterized in that: it is in containing the sample solution of aflatoxin B1, to add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, makes aflatoxin B1 fluorescent quenching.
Press such scheme, described for to make the fluorescence of aflatoxin B1 carry out abundant cancellation, first mix add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in the sample solution of aflatoxin B1 after, then in 37 ± 5 ℃, place at least 0.5h.
Press such scheme, also comprise according to following methods and measure fluorescent quenching rate: by the sample solution that contains aflatoxin B1 before not adding aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, prior to 365nm excitation wavelength, excite down and obtain its fluorescence emission spectrum, record the fluorescence intensity level of 440nm place (wavelength corresponding to fluorescence intensity maximum in fluorescence emission spectrum); And then add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 to carry out after cancellation, in 365nm excitation wavelength, excite down and obtain its fluorescence emission spectrum again, record the fluorescence intensity level of 440nm place (wavelength corresponding to fluorescence intensity maximum in fluorescence emission spectrum), according to following formula, calculate fluorescent quenching rate:
Fluorescent quenching rate=fluorescence added the difference of two fluorescence intensities at the 440nm place before and after aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation/the add fluorescence intensity level before aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation by aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation intensity level.
Press such scheme, while adding aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in said method in containing the sample of aflatoxin B1, make the concentration of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 reach the Cmin of the abundant cancellation of fluorescence that makes aflatoxin B1 in system, so that the fluorescence of aflatoxin B1 carries out abundant cancellation in sample.
Press such scheme, the implication of the aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 Cmin of the described abundant cancellation of fluorescence that makes aflatoxin B1 in system is: when in system, aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 is a certain concentration, increase along with this concentration, in system, the fluorescent quenching rate of aflatoxin B1 does not increase thereupon yet, and this concentration is the aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 Cmin of the abundant cancellation of fluorescence that makes aflatoxin B1 in system.Concrete preparation method is as follows: by the sample that contains aflatoxin B1 before not adding aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, prior to 365nm excitation wavelength, excite down and obtain its fluorescence emission spectrum, record the fluorescence intensity level of 440nm place (wavelength corresponding to fluorescence intensity maximum in fluorescence emission spectrum), then in above-mentioned aflatoxin B1 standard solution, add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, obtain the system of the malicious general purpose single clonal antibody of variable concentrations aspergillus flavus resisting, adopt as above condition to carry out fluorescent quenching, fluorescence intensity level after cancellation is compared with initial fluorescent intensity value, fluorescent quenching rate while obtaining variable concentrations aspergillus flavus resisting toxin B1 antibody, by comparing its fluorescent quenching rate, when aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 is a certain concentration, increase along with this concentration, in system, the fluorescent quenching rate of aflatoxin B1 does not increase thereupon yet, this concentration is the aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 Cmin of the abundant cancellation of fluorescence that makes aflatoxin B1 in system.
Aflatoxin B1 under state of nature can be sent fluorescence when excited by ultraviolet light (365nm), the wavelength that in fluorescence emission spectrum, fluorescence intensity maximum is corresponding is 440nm, and when being subject to ultraviolet light (365nm) again exciting after the specific aspergillus flavus resisting toxin of this aflatoxin B1 and its general purpose single clonal antibody 1C11 reacts, the fluorescence of aflatoxin B1 can the overwhelming majority be quenched.
Beneficial effect of the present invention:
Aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 can make aflatoxin B1 fluorescent quenching, and fluorescent quenching rate can reach 90%;
Simple to operate controlled;
Based on aspergillus flavus resisting toxin general purpose single clonal antibody, 1C11 can carry out the characteristic of cancellation to the fluorescence of aflatoxin B1, can enrich the bioanalysis to aflatoxin B1, the research means of detection.As the characteristic of aflatoxin B1 fluorescence being carried out to cancellation based on aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 can be carried out fast detecting etc. to aflatoxin B1 content in agricultural product.
Accompanying drawing explanation
Fig. 1 is the comparison of fluorescence intensity before and after the aflatoxin B1 standard solution of series concentration reacts with its aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, in figure: in two row corresponding to each concentration value: left column be the fluorescence intensity of each concentration aflatoxin B1 standard solution, and the right side is classified as and added aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 to react the fluorescence intensity after cancellation.
Fig. 2 is aflatoxin B1 concentration standard curve c
1(x
1, y
1), concentration range is 0.1～1ng/mL;
Fig. 3 is aflatoxin B1 concentration standard curve c
2(x
2, y
2), concentration range is 1～10ng/mL.
Embodiment
Following aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 used is that to adopt deposit number be that the method for reporting in the hybridoma cell strain 1C11 of the CCTCC NO:C201013 patent that is CN201010245095.5 according to number of patent application makes in advance, and concrete preparation method is as follows:
The BALB/c mouse of processing with freund 's incomplete adjuvant in advance with the hybridoma cell strain 1C11 injection that deposit number is CCTCC NO. C201013, collect the ascites of this mouse, adopt caprylic acid-ammonium antibody purification, concrete operations are: with double-deck Filter paper filtering mouse ascites, 4 ℃, the centrifugal 15min of 12000r/min, draw supernatant, gained ascites supernatant is mixed with the acetate buffer of 4 times of volumes, under stirring, slowly add caprylic acid, every milliliter of required caprylic acid volume of ascites is 33 μ L, mixed at room temperature 30min, 4 ℃ of standing 2h, then 4 ℃, the centrifugal 30min of 12000r/min, abandon precipitation, by the supernatant obtaining with after double-deck Filter paper filtering, the volumetric molar concentration that adds 1/10 filtrate volume is the phosphate buffer that 0.1mol/L and pH value are 7.4, with the sodium hydroxide solution of 2 mol/L, regulate the pH value to 7.4 of this mixed liquor, 4 ℃ of precoolings, slowly adding ammonium sulfate to ammonium sulfate final concentration is 0.277g/mL, 4 ℃ of standing 2h, then 4 ℃, the centrifugal 30min of 12000r/min, abandon supernatant, gained precipitation is resuspended with the 0.01mol/L phosphate buffer of former ascites volume 1/10, pack bag filter into, pure water is dialysed, the protein solution of fully having dialysed is put to-70 ℃ of refrigerator freezings, use afterwards freeze drier freeze-drying, collect freeze-dried powder, obtain the aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 that purifying is good, antibody is put in-20 ℃ of refrigerators standby,
Described acetate buffer is 0.29g sodium acetate, and 0.141mL acetic acid adds water and is settled to 100mL gained; The phosphate buffer of described 0.1mol/L is 0.8g sodium chloride, 0.29g disodium hydrogen phosphate, and 0.02g potassium chloride, potassium dihydrogen phosphate 0.02g, adds water and is settled to 100mL gained.
The aflatoxin B1 standard solution of preparation 10ng/mL, scan the fluorescence emission spectrum of this sample solution under 365nm excitation wavelength excites, record the fluorescence intensity level at 440nm place, in this sample solution, add again aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, mix, then in 37 ± 5 ℃, place 1h, make aflatoxin B1 fluorescent quenching, then in 365nm excitation wavelength, excite its fluorescence emission spectrum of lower acquisition, record the fluorescence intensity level at 440nm place, calculate fluorescent quenching rate, fluorescent quenching rate=fluorescence added the difference of two fluorescence intensities at the 440nm place before and after aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation/the add fluorescence intensity level before aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation by aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation intensity level.
Specifically in the aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 that adds different amounts makes system, the concentration of aspergillus flavus resisting toxin B1 antibody is respectively 0.1,0.5,5,10,20,30,40ng/mL, fluorescent quenching rate result is as follows respectively: when aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 0.1ng/mL, and the fluorescent quenching rate 31% of aflatoxin B1; When aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 0.5 ng/mL, cancellation rate is 37%; When aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 5ng/mL, cancellation rate is 49%; When aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 10ng/mL, cancellation rate is 78%; When aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 20ng/mL, cancellation rate is 91%, and in system aspergillus flavus resisting toxin general purpose single clonal antibody content continue to increase up to antibody concentration be 30 or during 40ng/mL, cancellation rate is still 91%.
Dilution phosphate buffer dilution for the aflatoxin B1 methyl alcohol storing solution that is 500ng/mL by concentration, configuration obtains the aflatoxin B1 standard solution (concentration is respectively 10,7.5,5,3.5,2,1.5,1,0.75,0.5,0.3,0.1 and 0 ng/mL) of series concentration, described phosphate-buffered formula of liquid is as follows: 0.8g sodium chloride, 0.29g disodium hydrogen phosphate, 0.02g potassium chloride, 0.02g potassium dihydrogen phosphate, adds water and is settled to 1000mL.The aflatoxin B1 standard solution 2mL that gets each series concentration excites under 365nm excitation wavelength, obtains the fluorescence emission spectrum of the aflatoxin B1 standard solution of each series concentration, and the fluorescence intensity level at recording wavelength 440nm place, is shown in Fig. 1.Then in the aflatoxin B1 standard solution of each concentration, add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, make the final concentration of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in each system be 20 μ g/mL, mix, place 1h, get each system solution 2mL, under 365nm excitation wavelength, excite, obtaining is respectively the fluorescence emission spectrum of system, get the fluorescence intensity level at maximum wavelength 440nm place, see Fig. 1.
Embodiment 3
Based on aspergillus flavus resisting toxin general purpose single clonal antibody, 1C11 carries out the characteristic of cancellation to aflatoxin B1 fluorescence, and the interpolation that can be used for Aflatoxin in Peanut byHigh B1 detects application experiment, and concrete application process is as follows:
I fluorescence is by relation curve c(x, the y of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation intensity and aflatoxin B1 concentration) foundation
(1) dilution phosphate buffer dilution for the aflatoxin B1 methyl alcohol storing solution that is 500ng/mL by concentration, configuration obtains the aflatoxin B1 standard solution (concentration is respectively 10,7.5,5,3.5,2,1.5,1,0.75,0.5,0.3,0.1 and 0 ng/mL) of series concentration, described phosphate-buffered formula of liquid is as follows: 0.8g sodium chloride, 0.29g disodium hydrogen phosphate, 0.02g potassium chloride, 0.02g potassium dihydrogen phosphate, adds water and is settled to 1000mL.The aflatoxin B1 standard solution 2mL that gets each series concentration excites under 365nm excitation wavelength, obtains the fluorescence emission spectrum of the aflatoxin B1 standard solution of each series concentration, and the fluorescence intensity level at recording wavelength 440nm place, is shown in Fig. 1.
(2) in the aflatoxin B1 standard solution of above-mentioned 10ng/mL, add respectively the different aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 that measure, (in system, the concentration of aspergillus flavus resisting toxin B1 antibody is respectively 0.1 to the system of acquisition variable concentrations aspergillus flavus resisting poison general purpose single clonal antibody 1C11, 0.5, 5, 10, 20, 30, 40ng/mL), adopt as above condition to carry out fluorescent quenching, fluorescence is compared with adding the fluorescence intensity level before aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation by aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation intensity level, obtain the fluorescent quenching rate of the aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 of variable concentrations.Concrete outcome: when aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 0.1ng/mL, the fluorescent quenching rate 31% of aflatoxin B1; When aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 0.5 ng/mL, cancellation rate is 37%; When aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 5ng/mL, cancellation rate is 49%; When aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 10ng/mL, cancellation rate is 78%; When aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 20ng/mL, cancellation rate is 91%, and in system, continue to increase up to antibody concentration be 30 or during 40ng/mL to aspergillus flavus resisting toxin general purpose single clonal antibody content, cancellation rate is still 91%, when aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 concentration is 20ng/mL in explanation system, can make the abundant cancellation of fluorescence of aflatoxin B1.Therefore in conjunction with cost, selecting the concentration of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in following system is 20ng/mL.
(3) in each aflatoxin B1 standard solution of step (1), add respectively aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, make the final concentration of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in each system be 20 μ g/mL, the hybridoma cell strain 1C11 that described aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 is CCTCC NO:C201013 by preserving number produces, mix, place 1h, get each system solution 2mL, under 365nm excitation wavelength, excite, obtaining is respectively the fluorescence emission spectrum of system, get the fluorescence intensity level at maximum wavelength 440nm place, see Fig. 1.
(4) scope that is 0.1-1ng/mL to aflatoxin B1 standard items concentration and the scope of 1-10ng/mL carry out respectively matching be piecewise fitting while obtaining small concentration fluorescence by the relation curve c of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation intensity and aflatoxin B1 concentration
1(x
1, y
1), curvilinear equation is: y
1=10
6x
1+ 8859, wherein: x
1aflatoxin B1 concentration, 0.1-1ng/mL, y
1for fluorescence added the difference of the 440nm place fluorescence intensity of aspergillus flavus resisting toxin B1 antibody 1C11 front and back by aspergillus flavus resisting toxin B1 antibody 1C11 cancellation intensity, see Fig. 2, and while obtaining large concentration fluorescence by the relation curve c of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation intensity and aflatoxin B1 concentration
2(x
2, y
2), curvilinear equation is: y
2=71535x
2+ 10
6, wherein: x
2aflatoxin B1 concentration, 1-10ng/mL, y
2for fluorescence added the difference of the 440nm place fluorescence intensity of aspergillus flavus resisting toxin B1 antibody 1C11 front and back by aspergillus flavus resisting toxin B1 antibody 1C11 cancellation intensity, see Fig. 3.
The interpolation of II Aflatoxin in Peanut byHigh B1 detects:
Take 5 grams of the peanut blank samples of milled, the methyl alcohol storing solution that adds 100 μ L aflatoxin B1, the interpolation concentration of aflatoxin B1 is 10 ng/g peanut blank samples, place half an hour, after methyl alcohol is evaporated completely, adding 15mL volumetric concentration is 80% methanol-water (sodium chloride that is 4% containing massfraction), under 50～60 ℃ of water-baths ultrasonic 10 minutes, quantitative filter paper filters, get filtrate 4mL and add 2mL sherwood oil, vortex 1 minute, stratification, take off a layer methanol-water 3mL, add 9 mL water, mix, it by the sample extracting solution of this dilution, with aperture, is the mixing membrane filtration of 0.45 μ m, get 10mL filtrate flow and cross aflatoxin B1 immune affinity column, with 1mL methanol-eluted fractions affinity column, collect meoh eluate, at 60 ℃, with nitrogen, dry up, with the above-mentioned dilution of 2mL, aflatoxin is hanged again, be sample solution, the fluorescence emission spectrum of scanning samples solution under 365nm excitation wavelength excites, again to add in sample solution aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 to aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 final concentration be 20 μ g/mL, mix, place after 1 hour, in 365nm excitation wavelength, excite its fluorescence emission spectrum of lower acquisition, record the fluorescence intensity level at 440nm place.
Testing result: by antibody cancellation intensity level, when testing sample solution fluorescence added to the large concentration of difference substitution of the 440nm place fluorescence intensity before and after aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 by antibody cancellation intensity level, fluorescence is by the relation curve c of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation intensity and aflatoxin B1 concentration according to system fluorescence after adding aspergillus flavus resisting toxin general purpose single clonal antibody 1C11
2(x
2, y
2) in (as Fig. 2-2), obtain the aflatoxin B1 concentration in sample solution, again in conjunction with the dilution situation in sample preparation process, the interpolation concentration that in processing procedure, the variation relation of concentration calculates the aflatoxin B1 in actual peanut sample is per sample 9.4 ng/g.
Based on aspergillus flavus resisting toxin general purpose single clonal antibody, 1C11 carries out the characteristic of cancellation to aflatoxin B1 fluorescence, can be used for the detection application experiment of aflatoxin B1 content in peanut sample, and concrete application process is as follows:
I fluorescence is by relation curve c(x, the y of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation intensity and aflatoxin B1 concentration) foundation as described in above-described embodiment 3.
The detection of II Aflatoxin in Peanut byHigh B1 content:
Take the peanut sample 1# to be measured of milled, 5 grams of 2# or 3#, adding 15mL volumetric concentration is 80% methanol-water (sodium chloride that is 4% containing massfraction), under 50～60 ℃ of water-baths ultrasonic 10 minutes, quantitative filter paper filters, get filtrate 4m L and add 2mL sherwood oil, vortex 1 minute, stratification, take off a layer methanol-water 3mL, add 9mL water, mix, it by the sample extracting solution of this dilution, with aperture, is the mixing membrane filtration of 0.45 μ m, get 10mL filtrate flow and cross the immune affinity column of aflatoxin B1, with 1mL methanol-eluted fractions affinity column, collect meoh eluate, at 60 ℃, with nitrogen, dry up, with the above-mentioned dilution of 2mL, aflatoxin is hanged again, be sample solution, the fluorescence emission spectrum of scanning samples solution under 365nm excitation wavelength excites, record the fluorescence intensity level at 440nm place, in sample solution, add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 again, to aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 final concentration be 20 μ g/mL, mix, place after 1 hour, in 365nm excitation wavelength, excite its fluorescence emission spectrum of lower acquisition, record the fluorescence intensity level at 440nm place.
Testing result: by the value of antibody cancellation, determine when the fluorescence of each sample solution added to the difference substitution small concentration of the 440nm fluorescence intensity before and after antibody by the value of antibody cancellation that fluorescence is by the relation curve c of aspergillus flavus resisting toxin general purpose single clonal antibody cancellation intensity and aflatoxin B1 concentration according to system fluorescence after adding aflatoxin b1 antibody
1(x
1, y
1) or during large concentration fluorescence by the relation curve c of aspergillus flavus resisting toxin general purpose single clonal antibody cancellation intensity and aflatoxin B1 concentration
2(x
2, y
2), obtain the aflatoxin B1 concentration in sample solution, again in conjunction with the dilution situation in sample preparation process, in processing procedure, the variation relation of concentration calculates the concentration of the aflatoxin B1 in peanut sample per sample, obtaining aflatoxin B1 concentration in the solution to be measured of peanut sample 1 is below detection limit 0.1ng/mL, in sample 1# aflatoxin B1 content for not detecting; In the solution to be measured of peanut sample 2, aflatoxin B1 concentration is 0.6ng/mL, and in sample 2#, aflatoxin B1 content is 2.16 μ g/kg; In the solution to be measured of peanut sample 3, aflatoxin B1 concentration is 3.2ng/m L, and in sample 3#, aflatoxin B1 content is 11.52 μ g/kg.Adopt national standard method GB/T 18979-2003 to detect above-mentioned peanut sample 1# simultaneously, 2# or 3#, its testing result is followed successively by: in peanut sample 1#, aflatoxin B1 does not detect, in peanut sample 2#, aflatoxin B1 content is 2.3 μ g/kg, and in peanut sample 3#, aflatoxin B1 content is 12.2 μ g/kg.In conjunction with above-mentioned detection, can find out: the testing result relative differences of above-mentioned two kinds of detection methods is all in 10%.
Claims (5)
1. the application of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in aflatoxin B1 fluorescent quenching, it is characterized in that: it is in containing the sample solution of aflatoxin B1, to add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, make aflatoxin B1 fluorescent quenching, wherein: the hybridoma cell strain 1C11 secretion that described aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 is CCTCC NO. C201013 by preserving number produces.
2. the application of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 according to claim 1 in aflatoxin B1 fluorescent quenching, it is characterized in that: described for to make the fluorescence of aflatoxin B1 carry out abundant cancellation, add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in the sample solution of aflatoxin B1 after, first mix, then in 37 ± 5 ℃, place at least 0.5h.
3. the application of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 according to claim 2 in aflatoxin B1 fluorescent quenching, it is characterized in that: also comprise according to following methods and measure fluorescent quenching rate: by the sample solution that contains aflatoxin B1 before not adding aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, prior to 365nm excitation wavelength, excite down and obtain its fluorescence emission spectrum, record the fluorescence intensity level at 440nm place; And then add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 to carry out after cancellation, then excite down and obtain its fluorescence emission spectrum in 365nm excitation wavelength, record the fluorescence intensity level at 440nm place, according to following formula, calculate fluorescent quenching rate:
Be fluorescence added the difference of two fluorescence intensities at the 440nm place before and after aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation/the add fluorescence intensity level before aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation by aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 cancellation intensity level to fluorescent quenching rate.
4. the application of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 according to claim 2 in aflatoxin B1 fluorescent quenching, it is characterized in that: while adding aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 in containing the sample of aflatoxin B1, make the concentration of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 reach the Cmin of the abundant cancellation of fluorescence that makes aflatoxin B1 in system, so that the fluorescence of aflatoxin B1 carries out abundant cancellation in sample.
5. the application of aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 according to claim 4 in aflatoxin B1 fluorescent quenching, it is characterized in that: described in to make the aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 Cmin of the abundant cancellation of fluorescence of aflatoxin B1 in system be to obtain according to following method: by the sample that contains aflatoxin B1 before not adding aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, prior to 365nm excitation wavelength, excite down and obtain its fluorescence emission spectrum, record the fluorescence intensity level at 440nm place, then in above-mentioned aflatoxin B1 standard solution, add aspergillus flavus resisting toxin general purpose single clonal antibody 1C11, obtain the system of the malicious general purpose single clonal antibody of variable concentrations aspergillus flavus resisting, adopt as above condition to carry out fluorescent quenching, fluorescence intensity level after cancellation is compared with initial fluorescent intensity value, fluorescent quenching rate while obtaining variable concentrations aspergillus flavus resisting toxin general purpose single gram antibody, by comparing its fluorescent quenching rate, when aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 is a certain concentration, increase along with this concentration, in system, the fluorescent quenching rate of aflatoxin B1 does not increase thereupon yet, this concentration is the aspergillus flavus resisting toxin general purpose single clonal antibody 1C11 Cmin of the abundant cancellation of fluorescence that makes aflatoxin B1 in system.
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Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1527942A (en) * | 2001-07-13 | 2004-09-08 | ���Ǿ� | Fluorescence polarizaton-based homogenous assay for aflatoxins |
| CN1588068A (en) * | 2004-09-07 | 2005-03-02 | 上海大学 | Detecting method for aflatoxins |
| CN1673746A (en) * | 2005-04-15 | 2005-09-28 | 中国农业科学院油料作物研究所 | Fast detecting method for aftatoxin B1 |
| CN2909243Y (en) * | 2006-06-20 | 2007-06-06 | 中国农业科学院油料作物研究所 | Quick investigating device of flavatin B |
| CN101109735A (en) * | 2007-05-14 | 2008-01-23 | 劲牌有限公司 | Fluorescence photometry for immune affinity column of aflatoxin in paddy |
| CN101949854A (en) * | 2010-09-14 | 2011-01-19 | 云南健牛生物科技有限公司 | Method for rapidly detecting content of aflatoxin |
| CN101993855A (en) * | 2010-08-05 | 2011-03-30 | 中国农业科学院油料作物研究所 | Hybridoma cell line 1C11 and anti-aflatoxin general monoclonal antibody generated by same as well as applications thereof |
| CN202101939U (en) * | 2011-05-17 | 2012-01-04 | 中国农业科学院油料作物研究所 | Measuring device for aflatoxin |
| JP2012177607A (en) * | 2011-02-25 | 2012-09-13 | National Agriculture & Food Research Organization | Method for detecting aflatoxin, device for detecting aflatoxin, and program |
| CN102747042A (en) * | 2012-04-20 | 2012-10-24 | 中国农业科学院油料作物研究所 | Hybridoma cell line 10G4 and anti-alfatoxin total (B1, B2, G1 and G2) monoclonal antibody produced by the same |
| CN102747043A (en) * | 2012-04-20 | 2012-10-24 | 中国农业科学院油料作物研究所 | Hybridoma cell line 3G1 and anti-alfatoxin B1 monoclonal antibody produced by the same |
-
2012
- 2012-11-30 CN CN201210501565.9A patent/CN103116024B/en not_active Expired - Fee Related
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1527942A (en) * | 2001-07-13 | 2004-09-08 | ���Ǿ� | Fluorescence polarizaton-based homogenous assay for aflatoxins |
| CN1588068A (en) * | 2004-09-07 | 2005-03-02 | 上海大学 | Detecting method for aflatoxins |
| CN1673746A (en) * | 2005-04-15 | 2005-09-28 | 中国农业科学院油料作物研究所 | Fast detecting method for aftatoxin B1 |
| CN2909243Y (en) * | 2006-06-20 | 2007-06-06 | 中国农业科学院油料作物研究所 | Quick investigating device of flavatin B |
| CN101109735A (en) * | 2007-05-14 | 2008-01-23 | 劲牌有限公司 | Fluorescence photometry for immune affinity column of aflatoxin in paddy |
| CN101993855A (en) * | 2010-08-05 | 2011-03-30 | 中国农业科学院油料作物研究所 | Hybridoma cell line 1C11 and anti-aflatoxin general monoclonal antibody generated by same as well as applications thereof |
| CN101949854A (en) * | 2010-09-14 | 2011-01-19 | 云南健牛生物科技有限公司 | Method for rapidly detecting content of aflatoxin |
| JP2012177607A (en) * | 2011-02-25 | 2012-09-13 | National Agriculture & Food Research Organization | Method for detecting aflatoxin, device for detecting aflatoxin, and program |
| CN202101939U (en) * | 2011-05-17 | 2012-01-04 | 中国农业科学院油料作物研究所 | Measuring device for aflatoxin |
| CN102747042A (en) * | 2012-04-20 | 2012-10-24 | 中国农业科学院油料作物研究所 | Hybridoma cell line 10G4 and anti-alfatoxin total (B1, B2, G1 and G2) monoclonal antibody produced by the same |
| CN102747043A (en) * | 2012-04-20 | 2012-10-24 | 中国农业科学院油料作物研究所 | Hybridoma cell line 3G1 and anti-alfatoxin B1 monoclonal antibody produced by the same |
Non-Patent Citations (3)
| Title |
|---|
| 粮油产品黄曲霉毒素B_1检测技术研究进展;李培武等;《中国油料作物学报》;20050630;第27卷(第02期);77-81 * |
| 高特异性黄曲霉毒素B_1单克隆抗体的制备及特性研究;肖智等;《中国油料作物学报》;20110228;第33卷(第01期);66-69 * |
| 黄曲霉毒素的检测;韩秀娣，等;《第四军医大学吉林军医学院学报》;20050630(第2期);5 快速荧光法 * |
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