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1、jupiter柱-用于蛋白质和多肽的提纯（The Jupiter column for the purification of proteins and peptides）The | laboratory laboratory safety management | | laboratory design | analysis method | analysis message board | recruitment | instrument industry training | | dispensing Chromatography Atomic Absorption |LIMS syst
2、em | voice room |Jupiter columnUsed for the analysis and purification of proteins, oligomers, and polymers2 pore size is 300? used for protein macromolecules (molecular weight greater than 15000), oligomers and polymers analysis;2 stable use within 3000 hours in the pH1.5-10 range;2 protein molecule
3、s were not altered during analysis and purification;2 have 5 10 15 the fixed phase is available;2 each 5µ column has a production test chart.Peak shape, high sharpness and efficiencyJupiterTMDesigned to analyze and purify proteins and macromolecules. The ultra pure silica (99.99%) and the dense
4、 bonded phase make the peaks become sharper and almost have no particular adsorption on the isolated proteins. The dense bonding phase and larger surface area of silica gel make JupiterTMThe column is able to withstand higher sample load. The sharp peak of protein peak and no destructive special abs
5、orption column makes JupiterTM become the best biological purification column analysis.PICTURE(P137, F1) peptide hormonesSpecifications: 250 * 4.6mmMobile phase: A:0.1% three trifluoroacetic acid aqueous solutionB:0.1% three trifluoroacetic acid soluble in acetonitrile / water (90:10)Gradient: a) 8m
6、in in A/B (90:10) to A/B (74:26) (B increases by 2% per minute)B) 6min in A/B (74:26) to A/B (70:30) (B increases by 0.57% per minute)Flow rate: 1.0mL/minTest: UV214nmSample: 1. 8- arginine oxytocin (cysteine - casein - Glutamine - asparagine - cysteine - Essence - Gan - Gan -NH2)2. vasopressin (cys
7、teine - casein - Glutamine - asparagine - cysteine - prolyl - Gan - -NH2)3., 4-, silk, -8-, light, oxytocin (cysteine - casein - light filament - asparagine - cysteine - clear - fine - sperm -NH2)4., oxytocin (cysteine - casein - Glutamine - asparagine - cysteine - prolyl - Gan -NH2)Stable in pH1.5-
8、10In addition JupiterTM special bonding process to obtain the high bonding rate and bonding symmetry, and dense bonding also makes the JupiterTM column has a service life of 3000 hours in the range of pH1.5-10, the broad adaptability pH value makes the JupiterTM column more easily eluted protein con
9、taining a biologically active agent in the mobile phase.Low three acetic acid conditionWhen using ESI to introduce sample components into a mass spectrometry system, the presence of three trifluoroacetic acid is actually a defect. Three acetic acid at regular working concentration (0.1%), because of
10、 its ability to form particle pairs as competing ions, makes the degree of ionization of the sample in the mass spectrometry system lower. Therefore, although the presence of three trifluoroacetic acid changes the shape of the peak, the sensitivity of the LC/ESI-MS system is obviously reduced. And t
11、he JupiterTM column,In three trifluoroacetic acid containing only 0.01% under the condition of separation of protein also can obtain very good shape.PICTURE(P138, F1) proteins operate under 0.1%, three trifluoroacetic acidColumn: Jupiter 5 mu C4 300?Specifications: 250 * 4.6mmItem: 00G-4167-E0Flow r
12、ate: 1.0mL/minMobile phase: A:0.1% three trifluoroacetic acid aqueous solutionB:0.1% three acetic acid soluble in acetonitrileGradient: A/B in 20min (75:25) to A/B (5:95)Test: UV214nmTemperature: 35 DEG CSample: 1. insulin2. trypsinogen3. whey protein4. myoglobin5. carbonic anhydraseFast LC/MC analy
13、sis2 lower analysis time, solvent usage, and column equilibration time can be reduced by about 70% in some applications.2 although the sample pass rate has increased by about 70%. However, retention is maintained with good sensitivity, resolution, and accuracy of mass spectrometric data.The followin
14、g is a comparison of protein and polypeptide samples using JupiterTM 5µ, 300? LC/MC 50 * 2 column and Jupiter 5 300? 250 * 2.0mm column.PICTURE(P138, F2) proteins operate under 0.1%, three trifluoroacetic acidColumn: JupiterTM 5 mu C4 300?Specifications: 250 * 4.6mmItem: 00G-4167-E0Flow rate: 1
15、.0mL/minMobile phase: A:0.01% three trifluoroacetic acid aqueous solutionB:0.01% three acetic acid soluble in acetonitrileGradient: A/B in 20min (75:25) to A/B (5:95)Test: UV214nmTemperature: 35 DEG CSample: 1. insulin2. trypsinogen3. whey protein4. myoglobin5. carbonic anhydrasePICTURE(P138, F3, 4)
16、 LC/MS (C18)Sample: 1. cytochrome C2. calf serum albumin3. myoglobinMass spectrometry: HP5989Scan range: 500-1800 in 1 secondA:Column: Jupiter 5 mu C18 300?Specifications: 250 * 2.0mmItem: 00G-4053-B0Flow rate: 0.2mL/minMobile phase: A:0.1% three trifluoroacetic acid aqueous solutionB:0.1% three ace
17、tic acid soluble in acetonitrileGradient: A/B in 60min (95:5) to A/B (5:95)B:Column: Jupiter 5 mu C18 300?Specifications: 50 * 2.0mmItem: 00G-4053-B0Flow rate: 0.2mL/minMobile phase: A:0.1% three trifluoroacetic acid aqueous solutionB:0.1% three acetic acid soluble in acetonitrileGradient: A/B in 45
18、min (95:5) to A/B (5:95)Saves 70% analysis time2 reduce solvent consumption and column cost;2, the use of fast gradient elution has a very good resolution;2 a good column for building a method.For example, the mechanism of large peptides by reversed-phase chromatography column, the first is the sepa
19、ration of adsorption and desorption, the organic content in the mobile phase most of the polypeptide has not reached a certain degree, are retained in the column; as long as the organic solvent concentration is reached, will be quickly washed off. Therefore, there is a dramatic effect of retention t
20、ime changes slightly on peptide eluting solvent composition, when using short columns when (50mm), hope that the analysis time is short, the special needs of the rapidly changing gradient elution to achieve the organic content required. The following is a comparison of the separation of polypeptides
21、 using long columns (250mm) and short columns (50mm).PICTURE(P139, F1) proteins were analyzed using Jupiter C18Column: Jupiter 5 mu C18 300?Test: UV220nmFlow rate: 1.0mL/minSample size: 5 LSample: 1. alkaline phosphate2. cyanocobalamine3. ribonucleic acid4. insulin5. iron (trans) protein6. trypsin i
22、nhibitorA:Specifications: 250 * 4.6mmItem: 00G-4053-E0Mobile phase: A:0.1% three trifluoroacetic acid aqueous solutionB:0.1% three acetic acid soluble in acetonitrileGradient: a) A/B in 7min (95:5) to A/B (74:26) (B reduced by 3% per minute)B) A/B in 3min (74:26) to A/B (66:34) (B reduced by 2.67% p
23、er minute)C) A/B in 10min (66:34) to A/B (46:54) (B reduced by 2% per minute)B:Specifications: 50 * 4.6mmItem: 00B-4053-E0Mobile phase: A:0.1% three trifluoroacetic acid aqueous solutionB:0.1% three acetic acid soluble in acetonitrileGradient: a) A/B in 1min (100:0) to A/B (80:20) (B reduced by 20%
24、per minute)B) A/B in 1.5min (80:20) to A/B (65:35) (B reduced by 10% per minute)C) A/B in 1.5min (65:35) to A/B (53.5:46.5) (B reduced by 7.67% per minute)(d) 2min in A/B (53.5:46.5) to A/B (53.5:46.5) (B constant)PICTURE(P139, F2) proteins were analyzed using Jupiter C4Column: Jupiter 5 mu C4 300?F
25、low rate: 1.0mL/minTest: UV220nmSample size: 5 LSample: 1. alkaline phosphate2. cyanocobalamine3. ribonucleic acid4. insulin5. iron (trans) protein6. trypsin inhibitorA:Specifications: 250 * 4.6mmItem: 00G-4167-E0Mobile phase: A:0.1% three trifluoroacetic acid aqueous solutionB:0.1% three acetic aci
26、d soluble in acetonitrileGradient: a) A/B in 7min (95:5) to A/B (74:26) (B reduced by 3% per minute)B) A/B in 3min (74:26) to A/B (66:34) (B reduced by 2.67% per minute)C) A/B in 10min (66:34) to A/B (46:54) (B reduced by 2% per minute)B:Specifications: 50 * 4.6mmItem: 00B-4167-E0Mobile phase: A:0.1
27、% three trifluoroacetic acid aqueous solutionB:0.1% three acetic acid soluble in acetonitrileGradient: a) A/B in 1min (100:0) to A/B (80:20) (B reduced by 20% per minute)B) A/B in 1.5min (80:20) to A/B (65:35) (B reduced by 10% per minute)C) A/B in 1.5min (65:35) to A/B (53.5:46.5) (B reduced by 7.6
28、7% per minute)(d) 2min in A/B (53.5:46.5) to A/B (53.5:46.5) (B constant)QAIn the whole production process of the column, the quality of the product is strictly tracked and guaranteed.Each 5µ column of JupiterTM has an analytical certificate. The certificate contains the contents of each batchJ
29、upiter column filling test results to ensure stability and reproducibility between columns and batches. Here are the test information for each of the 5 JupiterTM columns.L particle analysis- particle size- aperture- particle size distribution- specific surface areaTotal residual metal content of LL
30、bonding rateTotal carbon contentSurface coverage rateL factory test- stability- metal sensitivityHydrophobicity- Protein Standard Test- long term testing (pH stability)- scanning electron microscopy (measurement of surface smoothness and particle shape)ReproducibilityHere is a JupiterTM column spect
31、rum of cytochrome C isolated from different species for testing the reproducibility between different batches of products.PICTURE(P140, F1) proteinsColumn: JupiterTM 5 mu C18 300?Specifications: 250 * 4.6mmItem: 00G-4053-E0Mobile phase: A:0.1% three trifluoroacetic acid aqueous solutionB:0.1% three
32、acetic acid soluble in acetonitrileGradient: A/B in 15min (75:25) to A/B (45:55) (B increases by 2% per minute)Flow rate: 1.0mL/minTest: UV220nmSample: 1. horse cytochrome C2. ox cytochrome C3. dog cytochrome CCapillary column2 0.30mm, 0.50mm, 0.70mm inner diameter efficient glass lined column;2 hig
33、h sensitivity to trace sample analysis;2 fill with JupiterTM analysis column packing, easy to analyze the method of transplant application;2 is stable in LC/MS systems and suitable for a variety of gradient conditions;2 1/8 inch (3mm) outer sleeve tubes are of high strength and durability.In additio
34、n to the characteristics of the conventional size analysis column, the liquid chromatographic capillary column with Jupiter packing improved the sensitivity to trace sample analysis. Jupiter capillary column silica gel packing, high sphericity, smooth surface, to ensure that its column effect is ver
35、y high, and can be used for a long time. High purity silica gel and special bonding process makes the silica surface bonding is very compact, so Jupiter? Residual silanol groups in the capillary column is not sufficient to retain the interference and influence on the separation of the elution peak t
36、o a minimum.Semi prepared and prepared column2 the use of JupiterTM5 product bonding technology to produce 10 15 silica gel matrix filler;2 stuffing, good sphericity, smooth surface and high mechanical strength;2 uniform pore structure, high load;2 high pH stability, easy to column cleaningJupiter?
37、Column using the same process is bonded silica gel composite filler, particle size of 10 and 15 if, in the preparation of particle size fillers into other products (such as 3µ and 5;), during the separation process, will be aware of these changes.Less padding is neededThe filler density of the
38、JupiterTM column is very small, which means that only a small amount of filler is used to fill the column, and considerable operating costs can be saved for a large number of preparation systems.Long service lifeExcellent silicone and bonding technologies allow Jupiter columns to last longer and are
39、 used in the pH1.5-10 range. As long as the columns are cleaned in time, the use time of the columns can be increased.PICTURE(P142, F1) cationic peptide standardColumn: Jupiter 5 mu C18 300?Specifications: 250 * 4.6mmItem: 00G-4053-E0Flow rate: 1.0mL/minMobile phase: A:0.1% three trifluoroacetic aci
40、d aqueous solutionB:0.1% three acetic acid soluble in acetonitrileGradient: A/B in 20min (100:0) to A/B (60:40) (B increases by 2% per minute)Test: UV210nmSample: 1. +1, diethylene glycol - Gan - Gan - Gan - Gan - Gan - Gan - bright - Lai - amide2., +2, La - Lys - Gan - Gan - Gan - Gan - Gan - Gan -
41、 Lai - amide3., +3, La - Gan - C - C - bright - Lai - C - bright - Lai - Gan - Lai - amide4., +4, La - casein - C - bright - Lai - C - bright - Lai - Gan - Lai - amidePICTURE(P142, F2) proteinsColumn: Jupiter 5 mu C4 300?Specifications: 50 * 4.6mmItem: 00B-4167-E0Flow rate: 1.0mL/minMobile phase: A:
42、0.1% three trifluoroacetic acid aqueous solutionB:0.1% three acetic acid soluble in acetonitrileGradient: a) A/B in 1min (100:0) to A/B (80:20) (B reduced by 20% per minute)B) 1.A/B in 5min (80:20) to A/B (65:35) (B reduced by 10% per minute)C) A/B in 1.5min (65:35) to A/B (53.5:46.5) (B reduced by
43、7.67% per minute)(d) 2min in A/B (53.5:46.5) to A/B (53.5:46.5) (B constant)Test: UV220nmSample: 1. alkaline phosphate2. cyanocobalamine3. ribonucleic acid4. insulin5. iron (trans) protein6. trypsin inhibitorPICTURE(P142, F3) bioactive peptidesColumn: Jupiter 5 mu C18 300?Specifications: 250 * 4.6mm
44、Item: 00G-4167-E0Flow rate: 1.0mL/minMobile phase: A:20mM, KH2PO4, pH4.0B: acetonitrileGradient: a) 5min in A/B (95:5) to A/B (80:20)B) 13min in A/B (80:20) to A/B (60:40)Test: UV214nmSample: 1. (vascular) bradykinin2. neurotensin3. bombesin4.EledoisinPICTURE(P142, F4) genetic variants of insulinCol
45、umn: Jupiter 5 mu C18 300?Specifications: 250 * 4.6mmItem: 00G-4053-E0Flow rate: 1.0mL/minMobile phase: A:0.1% three trifluoroacetic acid aqueous solutionB:0.1% three acetic acid soluble in acetonitrileGradient: A/B in 20min (70:30) to A/B (68:32)Test: UV210nmSample: 1. bovine insulin2. human insuli
46、n3. porcine insulinPICTURE(P143, F1) transfer RNA fragmentsColumn: Jupiter 5 mu C4 300?Specifications: 250 * 4.6mmItem: 00G-4167-E0Flow rate: 1.0mL/minMobile phase: A:0.1M, K3PO4, soluble in 0.75% isopropyl alcohol, pH6.8B:1.0M (NH4) 2SO4+0.1MK3PO4 is soluble in 0.75% isopropyl alcohol, pH6.8Gradient: 20min, A/B (0:100) to A/B (70:30);10min A/B (70/30) to A/B (100:0);B co