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HALO Protein
HALO protein columns from AMT (Advanced Material Technology) are made of core-shell material and offer 400 Å and 1000 Å pore sizes for unrestricted pore access of large proteins. With these pore sizes and core-shell technology, ultra-fast and high-resolution separations can be achieved. The particles are available with a diameter of 2.7µm and 3.4µm and with ES-C18, C4 and diphenyl modifications.
Technical Data
| Phase | Modification | Particle size | Pore size | Endcapping | C-content | USP | Surface area | pH range | Tmax |
| HALO Protein ES-C18 | C18 | 2.7 µm | 1000 Å | + | 1.4 %C | L1 | 15 m²/g | 1.0-8.0 | 90°Ca |
| HALO Protein C4 | C4 | 2.7 µm | 1000 Å | + | 0.6 %C | L26 | 15 m²/g | 2.0-9.0 | 90°Ca |
| HALO Protein Diphenyl | Diphenlmethyl | 2.7 µm | 1000 Å | + | N/A | L11 | 15 m²/g | 2.0-9.0 | 90°Ca |
a Low pH; b High pH
Applications
Higher resolution with HALO 400 Ã… diphenyl compared to FPP 300 Ã… diphenyl
HALO 400 Ã… diphenyl outperforms an FPP 300 Ã… diphenyl column with higher resolution and 2.7 times lower back pressure. The 400 Ã… pores allow better access to the bound phase.
TEST CONDITIONS
- Columns:
HALO 400 Å Diphenyl, 3.4 μm, 2.1 x 150 mm
FPP 300 Å Diphenyl, 1.8 μm, 2.1 x 150 mm
- Flow rate: 0.2 mL/min
- Temperature: 60 °C
- Injection volume: 2 μL of 2 mg/mL denosumab
- Instrument: Shimadzu Nexera
- Detection: PDA at 220 nm
- Mobile Phase A: 88/10/2 Water/ACN/n-Propanol + 0.1% DFA
- Mobile Phase B: 70/20/10 n-Propanol/ACN/Water + 0.1% DFA
- Gradient: 18-28% B in 20 min
High temperature comparison of the HALO 1000 Ã… phases
Three bound phases (C4, ES-C18 and diphenyl) are now available on surface porous HALO 1000 Ã… particles. Each phase offers unique selectivity for mAbs, as shown in this high temperature separation of trastuzumab.
TEST CONDITIONS
- Columns:
HALO 1000 Å C4, 2.7 μm, 2.1 x 150 mm
HALO 1000 Å ES-C18, 2.7 μm, 2.1 x 150 mm
HALO 1000 Å Diphenyl, 2.7 μm, 2.1 x 150 mm
- Flow rate: 0.4 mL/min
- Temperature: 80 °C
- Injection volume: 2 μL with 2 mg/mL
- Sample: trastuzumab in water/0.1% TFA
- Instrument: Shimadzu Nexera
- Detection: PDA at 280 nm
- Mobile Phase A: Water/0.1% TFA
- Mobile Phase B: ACN/0.1% TFA
- Gradient:
32-40% B in 16 min (C4 und ES-C18)
32-38% B in 12 min (Diphenyl)
Comparison of HALO 1000 Å Diphenyl with the competitor product PolyPhenyl at 40 °C
HALO 1000 Å diphenyl shows improved resolution, retention and peak area compared to a competitor column of 450 Å SPPs with polyphenyl phase. The increased retention clearly shows the advantage of unhindered large pore access to the bound phase. The HALO diphenyl phase can also be operated at 40 °C without any loss of peak area.
TEST CONDITIONS
- Columns:
HALO 1000 Å Diphenyl, 2.7 μm, 2.1 x 150 mm
SPP 450 Å PolyPhenyl, 2.7 μm, 2.1 x 150 mm
- Flow rate: 0.4 mL/min
- Temperature: 40 °C
- Injection volume: 2 μL with 2 mg/mL
- Sample: trastuzumab in water/0.1% TFA
- Instrument: Shimadzu Nexera
- Detection: PDA at 280 nm
- Mobile Phase A: Water/0.1% TFA
- Mobile Phase B: ACN/0.1% TFA
- Gradient: 30-45% B in 15 min
Downloads
Brochures
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AMT HALO 1000Ã… C4 Protein Column for High Resolution Separation of a Monoclonal Antibody
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AMT HALO 1000Ã… Diphenyl Brochure
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AMT HALO® 1000Å Diphenyl Brochure
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AMT HALO® Applications Collection Brochure
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AMT HALO Bioclass Fused-Core Columns Brochure 2018
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AMT HALO Bioclass Brochure
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AMT HALO® Catalog
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AMT HALO® Catalog 2023
Applications
AMT HALO MZ-Apps Rp HALO 1000Ã… Brochure
AMT HALO MZ-Apps Rp Halo 1000Ã… C4 High Resolution Separation of a Monoclonal Antibody Application
Technical support
AMT HALO Bioanalysis Solutions Guidebook
The right column for you - we will be happy to support you individually
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