CN101698832A - Production of gluten protein monoclonal antibody and application of cell strain thereof - Google Patents

Abstract

The invention discloses a gliadin protein monoclonal antibody hybridoma cell strain and application thereof, which belong to the technical field of medical biology. The gliadin protein monoclonal antibody hybridoma cell strain is CCTCC-C200944. The invention has the advantages that: 1) chromosomes are stable and can secrete IgG2a antibodies; 2) the secreted antibodies have high titer; the supernate cultured directly can reach 1:100,000; 3) the content of antibodies in mouse ascites can reach 1mg/mL; the titer of the antibodies can reach 1: 1,000,000; and the antibodies can be purified by protein G; and 4) the secreted antibodies can be used for a western blot experiment (1:1,000).

Description

The production of gluten protein monoclonal antibody and the application of cell strain thereof

Technical field

The present invention relates to proteic monoclonal antibody hybridoma cell strain of gliadin and uses thereof, belong to the technical field of biological of food.

Background technology

According to FAO (1995) report, the food allergy more than 90% is to be caused by the common irritated food of egg, milk, fish, crustaceans, peanut, soybean, nuts and wheat 8 classes.Gluten protein is the particularly main anaphylactogen of wheat of cereal, causes that modal illness is a celiac disease---because the chronic intestinal malabsorption syndromes due to can not tolerating seitan (gluten).Symptom has obstinate diarrhoea, weight loss, anaemia, weak, tetany etc.Gliadin is the major cause that causes celiac disease.Gluten protein mainly is present in the wheat goods, as flour, biscuit, feed, contain wheat additive etc., but has or not the bran wheat on the market, aims at celiac patients or other special requirements, when to distinguish.

Gliadin (gliadin) also claims prolamine, water insoluble and dehydrated alcohol, but be dissolved in 70%～80% ethanol.This proteinoid of gliadin mainly is present in the plant seed, and as zein, gliadine etc., in ripe wheat seed, prolamine accounts for 40% of storage protein total amount, is the key protein of forming gluten.Be divided into four kinds of hypotypes, α-, β-, γ-and ω-prolamine, account for 25%, 30%, 30% and 15% of prolamine total amount respectively.

The method that detects the anaphylactogen gliadine has ELISA, radiation anaphylactogen adsorption experiment, countercurrent immunoelectrophoresis, histamine release test etc.ELISA detection sensitivity height, specificity and good reproducibility, but analysis time is long, commercialized degree is low.Radiation anaphylactogen adsorption experiment is highly sensitive, accuracy good, but human serum is had dependency, and that people IgE antibody accounts for serum proportion is little, and antibodies specific is also uncertain, and specific combination ability difference, therefore difficult stdn.Countercurrent immunoelectrophoresis is easy and simple to handle, quick, but resolving power is low, the too late ELISA of tolerance range.Histamine release assay sensitivity and specificity are higher, but whether sensitivity is fresh very relevant with food, and histamine is measured and must be finished within 24h after the blood drawing, causes application limit.According to the relative merits of the whole bag of tricks, comprehensive selection ELISA detects ovalbumin.

At present, existing on the market prolamine detection kit, but mostly be external production, price height, haul-cycle time are long.The prolamine ELISA detection kit of our development with the anti-prolamine monoclonal antibody of homemade hybridoma cell strain excretory wrapper sheet, adopts the double antibodies sandwich method, can qualitative or detection by quantitative prolamine.Detection sensitivity height, specificity are good, and price is lower than abroad, has started the precedent of domestic production prolamine detection kit.

The content of invention

The technical problem to be solved in the present invention is: a strain gliadin protein monoclonal antibody hybridoma cell strain and an excretory monoclonal antibody thereof is provided.

Another technical problem that the present invention will solve is: gliadin protein monoclonal antibody hybridoma cell strain and excretory monoclonal antibody thereof the purposes in field of medical examination is provided.

For achieving the above object, the present invention is by the following technical solutions:

Gliadin protein monoclonal antibody hybridoma cell strain, deposit number are CCTCC-C200944.

A kind of by hybridoma cell strain CCTCC-C200944 excretory monoclonal antibody.

Gliadin protein monoclonal antibody hybridoma cell strain is used to prepare the purposes of Gliadin monoclonal antibody.

A kind of purposes that is used to prepare the reagent that detects the Gliadin anaphylactogen by hybridoma cell strain CCTCC-C200944 excretory monoclonal antibody.

Gliadin protein monoclonal antibody hybridoma cell strain 2D8 is preserved in Chinese typical culture collection center, the address on June 17th, 2009: Wuhan University, deposit number is CCTCC-C200944, the classification called after of suggestion: hybridoma.

We adopt above-mentioned hybridoma to prepare gliadin albumen monoclonal antibody.This monoclonal antibody can be used for the content by the Gliadin anaphylactogen in the method test sample of the Elisa and the immune marking.

Advantage of the present invention is: 1) karyomit(e) is stable, IgG secretion 2a type antibody; 2) the secretory antibody height of tiring, directly culture supernatant can reach 1: 10 ten thousand; 3) mouse ascites can reach 1mg/mL; Tire and to reach 1: 100 ten thousand.Can use protein G purifying; 4) secretory antibody can be used for Elisa and immunoblotting (westernblot) experiment (1: 1000).

The invention will be further described below in conjunction with the drawings and specific embodiments; it is not limitation of the invention; according to prior art well known in the art; embodiments of the present invention are not limited to this; therefore all this areas according to the conduct of disclosure of invention institute be equal to replacement, all belong to protection scope of the present invention.

Fig. 1 is monoclonal antibody chromosome analysis result.

Embodiment

Embodiment 1: the preparation hybridoma

Material and source:

Antigen: gluten protein (gliadin, Sigma)

Animal: 6-8 week ABLB/c mouse in age (laboratory animal institute of the Chinese Academy of Medical Sciences)

Myeloma cell line: SP2/0 (Chinese Academy of Sciences's typical case's culture is preserved council's cell bank)

Fully and Freund (SIGMA company)

RPMI-1640 basic medium, complete RPMI-1640 substratum, HT substratum, HAT substratum (above reagent is all available from Gibico company)

Method:

Immune animal:

Employing mixes method, with isopyknic antigenic solution and adjuvant uniform mixing.

Initial immunity: every mouse makes a call to the every pin 0.2mL of 4 pins (the abundant place of oxter inguinal lymph nodes), uses complete freund adjuvant; Immunity for the second time after 21 days: the same but full freund adjuvant that toos many or too much for use of dosage needle number; Immunity for the third time after 14 days: the dosage needle number is the same but do not add adjuvant; Booster immunization after 10 days (be generally merge preceding 3 days): 50 μ g im iv; Get the spleen blood sampling after three days.

The separation of feeder cell:

Non-immune BALB/c mouse is plucked the eyeball bloodletting.The negative control sera of separation of serum during as antibody test.Mouse is soaked in 5min in 75% alcohol after drawing neck to put to death, expose peritonaeum in dissecting to tear after cutting off skin of abdomen after fixing on the platen, with cotton ball soaked in alcohol put on the skin repeatedly wipe away after, carefully cut off an osculum at peritonaeum, carefully be blown into the incomplete substratum of 5mL, collect abdomen towards liquid with custom; The back is washed repeatedly with the incomplete substratum of 5mL, and twice abdomen closes in the centrifuge tube towards the liquid merging, and the centrifugal 10min of 1000r/min abandons supernatant.Add HAT be cultured to 5mL with sedimentary cell resuspended and mixing, do cell counting, then according to the cell counting result, add the HAT substratum, making cell concn is 2 * 10 ⁵/ mL.Cell suspension is joined in 4 96 orifice plates, and every hole 100 μ L (about two) put 37 ℃ of 5%CO then ₂Incubator in cultivate.

The preparation of immune spleen cell:, pluck the eyeball bloodletting with the BALB/c mouse of immunity.The positive control serum of separation of serum during as antibody test.Mouse is soaked in 5min in 75% alcohol after drawing neck to put to death, after fixing on the dissection platen, raise left abdomen skin, can see spleen, change the eye scissors tweezer, in super clean bench, cut off peritonaeum with aseptic operation, take out spleen and place the plate that fills the incomplete substratum of 10mL, washing gently, and reticular tissue is on every side peelled off in carefulness.Spleen is moved in another plate that fills the incomplete substratum of 10mL,, make the incomplete substratum in the splenocyte immersion plate with plunger crush and grind spleen in 200 purpose copper mesh.With suction pipe piping and druming for several times, make single cell suspension.For removing the megalump in the splenocyte suspension, available 100 order steel copper mesh filter.The results splenocyte suspension, the centrifugal 10min of 1000r/min, full substratum centrifuge washing 1-2 time of toing many or too much for use is resuspended in cell the incomplete substratum mixing of 10mL then, gets above-mentioned suspension and counts.Usually every mouse can get 1 * 10 ⁸-2.5 * 10 ⁸Individual splenocyte.

Myeloma cell line:

The myeloma cell grows and went down to posterity with 1: 10 usually soon, (37 ℃, the 5%CO2 incubator, with the complete RPMI-1640 substratum (Gibico company) that contains 15%FBS, before the fusion, make most cells be in logarithmic phase, the cellular form homogeneous, cell degree of converging moderate (approximately 10-80%), the results myeloma cell, the centrifugal 10min of 1000r/min, full substratum centrifuge washing 1-2 time toos many or too much for use, then cell is resuspended in the incomplete substratum mixing of 10mL, gets above-mentioned suspension counting.

Cytogamy:

Splenocyte and myeloma cell SP/0 were mixed in the 50mL centrifuge tube by 4: 1, add incomplete substratum to 30mL.Abundant mixing; The centrifugal 5-10min of 1000r/min is with supernatant exhaustion as far as possible; At the bottom of touching fusion pipe on the palm, make sedimentation cell loose evenly; In 30 seconds, add 50%PEG (pH=8.0) 1mL that is preheated to 37 ℃ with the 1mL suction pipe, drip gently along tube wall limit edged; In 60 seconds, cell is all sucked in the suction pipe immediately then, static 30 seconds, in 30 seconds, cell is blown in the centrifuge tube then, in 5min, add the incomplete substratum that 30mL is preheated to 37 ℃ with suction pipe; The centrifugal 10min of 1000r/min adds the 40mLHAT nutrient solution after the supernatant discarded; Packing 96 porocyte culture plates; After 7-10 days with the HT substratum HAT substratum that swaps out; (available common perfect medium after the 14th day); Often observe the hybridoma growing state, treat that it grows to hole floorage 1/10 sucking-off supernatant confession antibody test when above.The method of antibody test is the Elisa method, as negative control, as positive serum, carries out the Elisa routine operation with immune mouse serum with myeloma cell's culture supernatant.

The clone cultivates hybridoma:

The hybridoma suspension that vegetation is to be cloned, with the HT substratum that contains 15% serum be diluted to every milliliter contain 2.5,15 with 3 kinds of different extent of dilution of 50 cells; Add 5 * 10 by every milliliter ⁴-1 * 10 ⁵Cell proportion adds respectively in above-mentioned hybridoma suspension and completes in the Tissue Culture Plate of feeder cell; One of every kind of hybridoma packing 96 orifice plate, each extent of dilution 32 hole, every hole amount is 0.1mL, every hole hybridoma number is respectively 0.5,3 and 10; 37 ℃, 7.5%CO ₂Moistening cultivation 7-10 days macroscopic clone occurs and can detect antibody; Early inverted microscope is observed down, marks and has only single clonal growth hole, goes supernatant to do antibody test; Get health figure and detect the exaggerative cultivation of positive hole moral cell, and frozen.

Select the high clone of the positive value of Elisa, the strong floorage of cell viability covers＞1/4 and is amplified to 24 orifice plates and carries out subcloning greater than 1/2 floorage when cell concentration, choose subclone 2-3 of 10-15 clone, chose in 14 days behind the subcloning the good mensuration Elisa of cell state to its positive value high to choose 5-10 subclone 2-3 individual increasing; Carry out subclone again with method, totally three times, 20 generations.

Hybridoma Cell Culture

Perfect medium culturing cell with 10%, 37 ℃, 5%CO ₂, changed a not good liquor every one day; Cell is big and bright, and big or small homogeneous is good.

Embodiment 2:Elisa method screening hybridoma

Main agents:

Coating buffer: carbonate buffer solution pH=9.6, NaHCO ₃2.93g; Adding distil water is to 1000mL.

Washing fluid and diluent: T:PBST pH=7.4:NaCl 8.0g; KH ₂PO ₄0.2g; NaHPO ₄12H ₂O2.9g; KCl 0.2g; Tween 0.5mL adding distil water is to 1000mL.

Confining liquid: 1%BSA (1g/100mL purchases SIGMA company)

4. colour developing liquid: TMB, every 1mLTMB adds the antioxidant NaS of 3 μ L 2mg/mL ₂O ₃.5H ₂O

A liquid: 0.2M Na ₂HPO ₄(28.4g/L) 25.7mL

0.1M citric acid (19.2g/L) 24.3mL

H ₂O ₂0.1%; Adding distil water 50mL

B liquid: TMB3.90g

Citric acid 10.52g

EDTA?1.86g

Glycerine 2000mL

DMSO 300mL, heating fusion back adding distil water is to 10000mL.

5. stop buffer (2M H ₂SO ₄): distilled water 178.3mL dropwise adds the vitriol oil (98%) 21.7mL.

Method:

Wrapper sheet: concentration 5 μ g/ holes, 4 ℃ are spent the night, and knockout plate washing 3-6 time pats dry;

Sealing: the every hole 100 μ L of 1%BSA, 37 ℃, 1 hour;

One is anti-: get every hole culture supernatant 100 μ L and add in the enzyme plate, 37 ℃ 1 hour, wash plate 3 times, 2min/ time, pat dry.

Two is anti-: the sheep anti-mouse igg thinning ratio that adds the HRP mark is 1: 3000 (a SIGMA company), every hole 100 μ L, 37 ℃ 1 hour, wash plate 3 times, 2min/ time, pat dry.

Colour developing: add A liquid 50 μ L/ holes earlier; Add B liquid 50 μ L/ holes again.Lucifuge 15min.

Stop: 50 μ L/ hole 2M H ₂SO ₄

Wavelength 450nm record analysis.

Three. the result

In the test of tiring with an Elisa left side after the fusion: merge the 1/10-1/4 that about about the 11 days newborn hybridomas in back reach floorage, in culture supernatant, can detect.

The selection of cell strain needs to consider: the amount of the antibody in the supernatant is subjected to multifactorially to influence, the amount of the ability of the secretory antibody of the quantity of hybridoma, hybridoma, the growth conditions of hybridoma, remaining splenocyte, supernatant liquor or the like factor.Therefore the selection of cell strain can not see singly that valence value gets height, comprehensive each side factor to select.This experimental selection 2D8 cell strain increases with frozen, and is preserved in Chinese typical culture collection center, address on June 17th, 2009: Wuhan University, deposit number is CCTCC-C200944, the classification called after of suggestion: hybridoma.

Embodiment 3: preparation monoclonal antibody in the animal body

Material and source

Mouse: the BALB/c mouse of growing up, age＞15 week, (laboratory animal institute of the Chinese Academy of Medical Sciences)

Body weight was respectively No. 1 30.0g before handled in the abdominal cavity, No. 2 29.1g

Handling the back body weight with whiteruss is respectively 31.2g, No. 2 30.6g

Cell strain: CCTCC-C200944.

Substratum is selected: 10% foetal calf serum perfect medium, (G1bico)

Method

Select the good SP2/0 cell of growth conditions to carry out amplification cultivation, the cell stand density is unsuitable excessive, and preferably not 80% of super floorage.Approximately every mouse needs 8 bottles of (25cm culturing bottle) cells, and every bottle contains 5 * 10 approximately ⁶Cell.

Handle in the abdominal cavity: every 0.7mL of whiteruss that the injection high pressure is crossed in the mouse peritoneal, handle back 10 days to 2 months all available.

The injection cell: collect the hybridoma of logarithmic phase, in 1-2 resuspension physiological saline of centrifuge washing, (concentration is 1 * 10 to adjust concentration ⁷-2 * 10 ⁷/ mL), every mouse intraperitoneal injection 0.8mL.

The inoculation back is observed: inoculation back 1-5 days, and two mouse do not have significant discomfort, and body weight increases (0.7g of Zheng family and 0.8g respectively) slightly since the 6th day obvious bulge of belly, and body weight increases 0.9g and 1.1g within 1 day on foot.Must extract ascites (3-5mL) during the mouse weight loss in a week afterwards, ascites 3000rpm5min room temperature centrifugal collecting cell injects in the mouse body of having handled well in the abdominal cavity can accelerate the ascitogenous time, the 3rd day visible abdominal tympanites after injection.

The processing of ascites: 3000rpm room temperature centrifugal collecting cell, at 4 ℃ of 12000r/min, 15min removes upper strata oil and lower floor's cellular constituent and other throw out, collects supernatant, measures antibody titer, packing ,-70 ℃ are frozen standby, or freeze-drying is preserved.After measured, mouse ascites can reach 0.6-1mg/mL; Tire and to reach 1: 360 ten thousand.

Purification process: Hitrip protein G is installed on the protein purification instrument, flow velocity is speed 1mL/min, use earlier the 10mL deionized water balance, use 50mL level pad (50m M Tris-Cl again, pH=8.0) balance affinity column, the ascites of getting behind the ammonium sulphate precipitation is used sample 5mL on the sample ring, runs flat with level pad balance to baseline again, use elutriant (the 0.1M glycine pH=5.0) wash-out of 10mL again; Peak and elution peak are passed in collection, and elution peak transfers to the pH of elutriant about 7.2 rapidly with neutralizer (1M Tris-Cl pH=8.0).Each pipe of collecting is carried out SDS-PAGE gel electrophoresis analysis purity and concentration difference, select concentration and purity high.Concentrate 10000rpm, 4 ℃, 20min with the ultrafiltration pipe.(preservation of antibody: the BSA that adds final concentration and be 0.1% sodium azide and 1% is in-70 ℃ of prolonged preservation.)

Embodiment 4: in vitro method obtains monoclonal antibody

One, selects to carry out vitro culture, at first with containing 10% perfect medium culturing cell through the 2D8 cell strain of three subclonings.The 2D8 cell strain growth is active, can tolerate this process, but needs change every day liquid, stablely cultivates liquid nitrogen cryopreservation after 5 generations.

Two. purifying: utilize the protein g affinity chromatography method.: Hitrip protein G is installed on the protein purification instrument, flow velocity is speed 1mL/min, use earlier the 10mL deionized water balance, use 50mL level pad (50mM Tris-Cl again, pH=8.0) balance affinity column, the ascites of getting behind the ammonium sulphate precipitation is used sample 5mL on the sample ring, runs flat with level pad balance to baseline again, use elutriant (the 0.1M glycine pH=5.0) wash-out of 10mL again; Peak and elution peak are passed in collection, and elution peak transfers to the pH of elutriant about 7.2 rapidly with neutralizer (1M Tris-Cl pH=8.0).Each pipe of collecting is carried out SDS-PAGE gel electrophoresis analysis purity and concentration difference, select concentration and purity high.Concentrate 10000rpm, 4 ℃, 20min with the ultrafiltration pipe.(preservation of antibody: the BSA that adds final concentration and be 0.1% sodium azide and 1% is in-70 ℃ of prolonged preservation.)

Embodiment 6: the evaluation of antibody characteristic

The chromosome analysis of hybridoma

Method:

48-36h goes down to posterity hybridoma before adding colchicine.In culturing bottle, add colchicine (100 μ g/mL, degerming ,-20 ℃ of preservations), make ultimate density be 0.1-0.4 μ g/mL (if use instead Omaine then ultimate density be 0.02-0.05 μ g/mL); Continue to cultivate 4-6 hour, blow and beat cell then, move in the centrifuge tube, 1000r/min 10min abandons supernatant.Adding has been preheating to 37 ℃ 0.075mol/L KCL solution 5mL, sedimentation cell is suspended and mixing 37 ℃ of waters 15-20min.Stationary liquid (methyl alcohol mixes with Glacial acetic acid at 3: the 1) 1mL that in suspension, adds new preparation, mixing, 1000r/min10min then, abandoning supernatant: the purpose of this step is that cell surface is slightly fixing, the agglomerating money of cytoadherence after can preventing to fix.Add stationary liquid 5mL, with cell suspension and mixing, the static 20-30min of room temperature, 1000r/min10min then, abandoning supernatant; Repeat to do operation once; Add the 5mL stationary liquid,, seal up the mouth of pipe, put 4 ℃ and spend the night cell suspension and mixing thereafter.Take out centrifuge tube, 1000r/min 5min, the gentle aspiration supernatant liquor, stay the 0.5-1mL stationary liquid according to cell pack,, draw cell suspension 1-2 and drip with cell suspension and after being mixed, drop on the wave carrier piece that just from frozen water, takes out, dispel with mouth, and on flame, pass through to make cell be tiled in seasoning on the wave carrier piece for several times.10%Giemsa dye liquor dyeing 10-20min with new preparation draws dye liquor with tap water, seasoning then.

Microscopy: the selective staining body is scattered, and zero lap does not have the cell that scatters and wakes up with a start observation analysis.Every part of sample should be counted 100 complete metaphase nucleus cells, and notes observing whether marker chromosome is arranged.

The result: karyomit(e) is stable, sees Fig. 1.

Embodiment 7: the evaluation of monoclonal antibody heavy chain immunoglobulin and light chain type

One. material

1. coating buffer: carbonate buffer solution pH=9.6, Na ₂CO ₃1.59g; NaHCO ₃2.93g; First distilled water is to 1000mL.

2. washing fluid and diluent: T:PBST pH=7.4:NaCl 8.0g; KH ₂PO ₄0.2g; NaHPO ₄12H ₂O 2.9g; KCl 0.2g; Tween 0.5mL adding distil water is to 1000mL.

3. confining liquid: 1%BSA (Biochrom AG company)

4. colour developing liquid: TMB, every 1mLTMB adds the antioxidant NaS of 3 μ L 2mg/mL ₂O ₃.5H ₂O

A liquid: 0.2M Na ₂HPO ₄(28.4g/L) 25.7mL

0.1M citric acid (19.2g/L) 24.3mL

H ₂O ₂0.1%; Adding distil water 50mL

B liquid: TMB3.90g

Citric acid 10.52g

EDTA?1.86g

Glycerine 2000mL

DMSO 300mL, heating fusion back adding distil water is to 10000mL.

5. stop buffer (2M H ₂SO ₄): distilled water 178.3mL dropwise adds the vitriol oil (98%) 21.7mL.

Two methods:

(1)

1. wrapper sheet: concentration 50 μ g/mL, 4 ℃ are spent the night, and knockout plate washing 3-6 time pats dry;

2. sealing: 1%BSA (Biochrom AG company) sealed 1 hour in 37 ℃;

3. one is anti-: get every hole culture supernatant 100 μ L and add in the enzyme plate, 37 ℃ 1 hour, wash plate 3 times, 2min/ time, pat dry.

4. antibody subtype: the sheep anti-mouse antibody hypotype thinning ratio of no HRP mark is 1: 6000 (a SIGMA company), 37 ℃ 1 hour, wash plate 3 times, 2min/ time, pat dry.

4. two is anti-: the sheep anti-mouse igg thinning ratio of HRP mark is 1: 3000 (a SIGMA company), 37 ℃ 1 hour, wash plate 3 times, 2min/ time, pat dry.

5. colour developing: add A liquid 50/ hole earlier; Add B liquid 50/ hole again, 37 ℃ of 10min.

6. stop: 50 μ L/ hole 2M H ₂SO ₄

7. wavelength 450nm record analysis.

1. result: monoclonal antibody is an IgG2a type antibody.

(2) antibodies specific detects:

1. method: adopt conventional WesternBlot trace method (immunoblotting) to measure antibody purity, one anti-is Gliadin monoclonal antibody (behind the ascites purifying), two anti-are sheep anti-mouse igg/HRP (middle China fir Golden Bridge Bioisystech Co., Ltd), ECL test kit (Puli's lema gene technology company limited)

2. result:, almost do not have the specific reaction band to form with the many resistive connections of LRP16 high specificity relatively really.

(3) antibody titer is measured:

1. method: conventional ELISA method

2. reagent: with ELISA method screening hybridoma

3. result: the supernatant that nail is cultivated can reach 1: 20 ten thousand; Mouse ascites can reach 0.6-1mg/mL; Tire and to reach 1: 100 ten thousand to 1: 360 ten thousand.

Above-mentioned LRP16 monoclonal antibody is suitable for westernblot and immunohistochemical study uses, and the result shows this antibodies specific height, and background is clean.

Claims (1)

1. gliadin protein monoclonal antibody hybridoma cell strain CCTCC-C200944

A kind of by hybridoma cell strain CCTCC-C200944 excretory monoclonal antibody.

Gliadin protein monoclonal antibody hybridoma cell strain is used to prepare the purposes of Gliadin monoclonal antibody.

Gliadin protein monoclonal antibody hybridoma cell strain is used to prepare the purposes of the reagent that detects the Gliadin anaphylactogen.

A kind of purposes that is used to prepare the reagent that detects the Gliadin anaphylactogen by hybridoma cell strain CCTCC-C200944 excretory monoclonal antibody.

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