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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 Å diphenyl compared to FPP 300 Å diphenyl
HALO 400 Å diphenyl outperforms an FPP 300 Å diphenyl column with higher resolution and 2.7 times lower back pressure. The 400 Å 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 Å phases
Three bound phases (C4, ES-C18 and diphenyl) are now available on surface porous HALO 1000 Å 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 1000Å C4 Protein Column for High Resolution Separation of a Monoclonal Antibody
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AMT HALO 1000Å 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 1000Å Brochure
AMT HALO MZ-Apps Rp Halo 1000Å 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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