The Biozen dSEC columns are packed with low pore volume silica coupled with a proprietary hydrophilic diol-type bonded surface chemistry that prevents the silica surface from interacting with protein samples.
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Biozen Biologics LC Columns
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Explore Biozen LC Portfolio
Elevate your protein analysis with precision and versatility using Biozen's advanced LC columns. Optimize your sensitive HPLC analysis for superior results.
Explore Biozen LC Portfolio
With a new titanium BioTiâ„¢ biocompatible hardware to minimize priming, four particle platforms for optimal versatility and nine particle chemistries to maximize selectivity and sensitivity, Biozen UHPLC/HPLC columns for protein analysis are seamlessly designed to bring peace of mind to your analysis of biologics through:
Biocompatible Flow Path with BioTiâ„¢ Hardware
Keep your mind at ease knowing that we’ve minimized the need for priming with a new titanium infused biocompatible hardware and frit that doesn’t interfere with protein or peptide integrity!
A: UHPLC Pressure Rating Allows for more experimental design space; room to increase flow rates for increased throughput |
B: Better Controlled ID Versus PEEK Hardware Improved column-to-column performance, providing more consistency and easier method validation |
C: Biocompatible Titanium Liner |
D: Improved Recovery and Peak Shape Better column-to-column consistency, ease of use, and troubleshooting |
E: Strong Stainless Steel Walls Help Prevent Movement Under Pressure |
F: Biocompatible Titanium Frit |
4 Advanced Particle Platforms
All four of the Biozen particle platforms were individually designed and built by Phenomenex to take advantage of integral levels of performance, ruggedness, and reproducibility for protein characterization applications. Individually, each platform differs in the proprietary processing techniques used to control particle size and morphology.
Pore Controlled Technology
Monosized Polymeric
Non-Porous
Core-Shell Technology
Thermally Modified
Fully Porous
Pore Controlled
Technology
Monosized Polymeric Non-Porous
Meticulously controlled monosized particle technology secures incredible particle consistency that leads to improved and reliable efficiency. This innovative non-porous particle serves as the perfect backbone for complex ion-exchange chemistries.
Core-Shell Technology
Using sol-gel processing techniques that incorporate nano structuring technology, a durable, homogeneous porous shell is grown on a solid silica core. This highly optimized process combined with industry leading column packing technology produces highly reproducible columns that generate extremely high efficiencies and sensitivity.
Thermally Modified Fully Porous
Through a proprietary thermal processing series of steps, we eliminate micropores and further improve consistency, column efficiency, inertness, ruggedness, and reproducibility.
11 Particle Chemistries
Oligonucleotide Analysis
The Biozen Oligo LC column brings a unique combination of improved chromatographic efficiency from its core-shell morphology and high pH stability necessary for oligonucleotide characterization. Biozen Oligo is packed in a unique bio-inert titanium hardware developed to mitigate non-specific interactions that lead to analyte loss, distortions in peak shape and carry over. Confidently maximize resolution and reproducibility throughout the most demanding of method extremes with Biozen Oligo.
LC-MS Analysis of siRNA using BioTi UHPLC Hardware
Bio-inert hardware gives clear separation of the sense and anti-sense strand. View application>>
LC columns
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Columns:
Biozen 2.6 μm Oligo ( BioTi™)
Clarity 2.6 μm Oligo XT (stainless steel) |
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Dimensions
100 x 2.1 mm
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Part No.:
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Mobile Phase:
A: A: 4 mM Triethylamine in Water + 12.5 mM
Hexafluoro 2 propanol
B: B: 4 mM Triethylamine in Methanol + 12.5 mM Hexafluoro 2 propanol |
vs
LC-MS Analysis of siRNA using Stainless Steel UHPLC Hardware
Non-specific interactions were observed using stainless steel hardware resulting in bimodal peak for late eluting anti-sense strand.
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Gradient:
Time (min)
0
2
16
16.1
20
20.1
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% B
5
5
30
95
95
5
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Flow Rate:
0.3 mL/min
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Temperature:
55 °C
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Instrument:
Shimadzu® LC 20A Prominence®
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Detection:
TOF-MS
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Detector:
SCIEX® TripleTOF ® 6600
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Peptide Mapping
Digested mAbs or ADCs typically include a large body of compounds which are crucial to understanding post translation modifications. So we designed two Biozen Peptide columns to offer uniquely different selectivity profiles for these types of bioseparations. Each allows for fast and effective elution windows by utilizing either high efficiency core-shell or thermally modified fully porous particles to gain sharper peaks, better peak capacities, and overall higher sensitivity.
Trastuzumab Biosimilar Peptide Map
Infliximab Biosimilar Peptide Map
Conditions for all columns:
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Column:
 Biozen 1.6 µm Peptide PS-C18 Biozen 2.6 µm Peptide XB-C18 |
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Dimensions
150 x 2.1 mm
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Part No.:
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Mobile Phase:
A: 0.1 % Formic Acid in Water
B: 0.1 % Formic Acid in Acetonitrile |
Gradient:
Time (min)
0
0.5
50
55
56
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1
1
50
50
95 |
Flow Rate:
0.3 mL/min |
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Temperature:
40 °C |
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Detection:
QTOF (SCIEX® X500B) |
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With protein aggregate often at very low levels (<0.1% by peak area compared to monomer) and fragment separation a requirement, adequate resolution and peak shape have become even more crucial method outcomes. To address this need, the robust set of Biozen SEC columns were developed with a combination of UHPLC efficiency and higher sensitivity, to drive resolution and identification of even lower level targets. Biozen dSEC-7 Brochure>> Biozen dSEC-2 Brochure>>
LC Conditions:
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Column:
Biozen 3 µm dSEC-7
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Dimensions
150 x 4.6 mm
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Part No.:
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Mobile Phase:
200 Potassium Phosphate +
250 mM KCl, pH 6.2 |
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Part No.:
00H-4787-E0
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Mobile Phase:
20 mM Sodium Phosphate, pH 6.6 + 350 mM Potassium Chloride
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Flow Rate:
350 µL/min (Isocratic)
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Injection Volume:
 0.5 µL 1 µL 2 µL |
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Temperature:
25 °C
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Instrument:
Waters® ACQUITY® H-Class
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Detection:
FLR - Ex 280 nm, Em 350 nm
Sampling Rate: 40 Hz |
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Sample:
1. AAV2-CAG-GFP, 2E13 vg/mL
2. AAV5-CMV-GFP, 2E13 vg/mL 3. AAV8-CMV-GFP, 2E13 vg/mL 4. AAV9-CMV-GFP, 2E13 vg/mL |
Conditions for all columns:
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Column:
Biozen 1.8 µm dSEC-2, 200  Å
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Dimensions
300 x 4.6 mm
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Part No.:
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Mobile Phase:
200 Potassium Phosphate +
250 mM KCl, pH 6.2 |
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Part No.:
00H-4787-E0
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Flow Rate:
0.35 mL/min
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Injection Volume:
10 µL
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Temperature:
25 °C
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Detection:
UV @ 280 nm
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Sample:
Various,10 mg/mL
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Biozen WCX was crafted to consistently decipher between native protein variants that arise from PTMs within a therapeutics creation and development. The linear polycarboxylate chains grafted to monosized non porous polymeric particles, envelop and separate proteins from acidic and basic protein variants. With such a highly tuned and controlled manufacturing process, Biozen WCX HPLC columns affords scientists a way to reproducibly characterize heterogeneity while taking advantage of excellent recovery through high particle inertness and bioinert titanium BioTiâ„¢ column hardware. View Application Note>>
Trastuzumab (MES Salt Gradient)
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Column:
Biozen 6 μm WCX
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Dimensions
250 x 4.6 mm
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Part No.:
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Mobile Phase:
A: 20 mM MES (pH 5.6)
B: 20 mM MES + 300 mM NaCl (pH 5.6) |
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Gradient:
Time (min)
0
1
31
31.1
34
35
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% B
15
15
45
100
100
15
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Flow Rate:
1 mL/min
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Temperature:
30 °C
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Detection:
UV @ 280 nm
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Sample:
Trastuzumab
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Trastuzumab (pH Gradient Buffer)
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Column:
Biozen 6 μm WCX
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Dimensions
250 x 4.6 mm
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Part No.:
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Mobile Phase:
A: CX -1 (pH 5.6) pH Gradient Buffer*
B: CX -1 (pH 10.2) pH Gradient Buffer*
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Gradient:
Time (min)
0
1
21
23
24
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% B
0
0
100
100
0
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Flow Rate:
1 mL/min
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Temperature:
30 °C
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Detection:
UV @ 280 nm
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Sample:
Trastuzumab
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* From Thermo Fisher Scientific® Inc.
Intact and Subunit Analysis
Impurity profiling and characterization of intact biologics and fragments is a challenging undertaking because of the need to identify very small differences between variants. Both Biozen Intact columns. contain skillfully manufactured large pore core-shell particles that provide narrower, taller peaks in conjunction with higher resolution between the target mAb HC/LC, Fc/Fab, or isoforms.
Intact Trastuzumab at 70, 80, and 90 °C
Conditions for all columns:
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Column:
Biozen 3.6 µm Intact XB-C8
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Dimensions
150 x 2.1 mm
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Part No.:
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Mobile Phase:
A: 0.1 % TFA in Water
B: 0.1 % TFA in Acetonitrile
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Gradient:
Time (min)
0
1
13
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% B
20
20
25
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Flow Rate:
0.5 mL/min
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Temperature:
 70 °C 80 °C 90 °C |
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Detection:
UV @ 280 nm
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Sample:
1. Trastuzumab
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Infliximab F(ab)2
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Column:
Biozen 3.6 µm Intact XB-C8
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Dimensions
150 x 2.1 mm
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Part No.:
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Mobile Phase:
A: 0.1 % TFA in Water
B: 0.1 % TFA in Acetonitrile
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Gradient:
Time (min)
0
1
13
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% B
20
20
60
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Flow Rate:
0.5 mL/min
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Temperature:
80 °
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Detection:
UV @ 280 nm
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Sample:
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Cetuximab
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Column:
Biozen 3.6 µm Intact XB-C8
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Dimensions
150 x 2.1 mm
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Part No.:
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Mobile Phase:
A: 0.1 % TFA in Water
B: 0.1 % TFA in Acetonitrile
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Gradient:
Time (min)
0
1
13
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% B
20
20
45
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Flow Rate:
0.5 mL/min
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Temperature:
80 °
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Detection:
UV @ 280 nm
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Sample:
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Intact Mass Analysis
Intact mass can give indications not only of relative abundance of glycoforms , but also stability as degraded mAbs will not give good charge envelope by ESI-MS. Intact Mass with a high resolution MS to identify PTMs, especially relative abundance of glycoforms , combines extremely well with the fast run times and tight peak shapes provided by the Biozen Intact XB-C8 and Biozen WidePore C4.
Intact Mass of Trastuzumab Biosimilar using a Biozen Intact XB-C8 and SCIEX® X500B
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Column:
Biozen 3.6 µm Intact XB-C8
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Dimensions
150 x 2.1 mm
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Part No.:
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Mobile Phase:
A: 0.1 % Formic Acid in Water
B: 0.1 % Formic Acid in Acetonitrile /
Isopropyl alcohol (50:50
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Gradient:
Time (min)
2.5
10
110.1
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% B2
0
65
95
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Flow Rate:
0.3 mL/min
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Temperature:
90 °
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Detection:
QTOF (SCIEX X500B)
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Sample:
Trastuzumab
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Drug Antibody Ratio (DAR)
With a direct effect on efficacy and safety, conjugation for each ADC must be well understood. The Biozen Intact XB-C8 provides an excellent vehicle for determining drug load distribution and DAR for ADCs. Its large pore size allows intact ADCs to interact with a moderately retentive stationary phase while the core-shell particle supplies increased efficiency to deliver the required resolution between ADC species with differing drug loads.
Herceptin—vcMMAE using Biozen 3.6 µm Intact XB-C8
Herceptin—mcMMAF using Biozen 3.6 µm Intact XB-C8
Peptide Quantitation
When quantitating signature peptides from biological matrices, you need sharp peak shape and sufficient retention of hydrophilic peptides to prevent any signal loss from matrix suppression regions. Both Biozen Peptide columns were developed to deliver excellent selectivity for even closely related peptides. Additionally, they build on this body of valuable characteristics with unique ways of delivering sharper peak shape for basic peptides; Biozen Peptide XB-C18 blocks secondary surface interactions via isobutyl side chains, while the Biozen Peptide PS-C18 contains a positively charged weak base that repels other basic species.
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Column:
Biozen 3 µm Peptide PS-C18
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Dimensions
50 x 2.1 mm
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Part No.:
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Mobile Phase:
A: 0.1 % Formic Acid in Water
B: 0.1 % Formic Acid in Acetonitrile |
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Gradient:
Time (min)
0
1
4.5
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%B
3
3
25
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| Flow Rate:
0.5 mL/min
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| Temperature:
22 °C |
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| LC System:
ExionLCâ„¢ AD HPLC |
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Detection:
MS/MS
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Detector:
SCIEX QTRAP® 5500
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Flow Rate:
1.85 mL/min
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Sample:
As noted
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Glycan Analysis
The unique selectivity of the Biozen Glycan was designed to provide higher order separations of released and labeled glycans. With a 2.6 μm core-shell particle size, customers using either HPLC or UHPLC systems can draw upon a high efficiency Biozen Glycan particle run at higher linear velocities to easily provide sharper peak shapes and faster elution windows, without high UHPLC pressures. Under HILIC-FLR or HILIC-MS conditions, the Biozen Glycan excels with increased polar retention and selectivity.
Intact Mass of Trastuzumab Biosimilar using a Biozen Intact XB-C8 and SCIEX® X500B
Conditions for all columns:
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Column:
Biozen 2.6 µm Glycan
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Dimensions
150 x 2.1 mm
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Part No.:
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Mobile Phase:
A: 100 mM Ammonium Formate, pH 4.5
B: Acetonitrile |
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Gradient:
Time (min)
0
10
24
38.5
38.6
40.6 40.7 48 |
% B
78
74.5 72 55.9 40 40 78 78 |
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Flow Rate:
0.5 mL/min
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Temperature:
50 °C
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Detection:
FLD ex/em 285/345 nm
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Sample:
As noted
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Phenomenex is a technology leader committed to developing novel analytical chemistry solutions that solve the separation and purification challenges of researchers worldwide.
