Multiple Chemistries Provide Better Selectivity
and More Purification Options
MEP HyperCel™ Sorbent
- Stability of sorbent and ligand allows up to 200 cycles of purification
- Broad species and isotype binding capabilities for antibodies
- Elution at milder pH as compared to conventional antibody purification with Protein A
- Ligand structure and density of MEP HyperCel sorbent provides effective binding in the absence of lyotropic agents or salts
HEA and PPA HyperCel Sorbents
- HyperCel cellulose bead provides high porosity, chemical stability, and low non-specific interaction
- Binding based on hydrophobic interactions and elution on the basis of electrostatic repulsion
- Binding typically at physiological pH with no need to use high salt, unlike conventional hydrophobic interaction chromatography (HIC)
- HEA and PPA HyperCel sorbents offer effective discrimination of proteins having similar or very close isoelectric points
MEP HyperCel Sorbent
- Antibody capture and purification
- Purification of Fc fusion proteins and Ab-like molecules
- Easily screen multiple mixed-mode chemistries
HEA and PPA HyperCel Sorbent
- Direct hydrophobic capture and purification
- Compatible upstream and downstream of ion exchange and other chemistries for enhanced purification
- Easily screen multiple mixed-mode chemistries
Materials of Construction
- Column Housing, Cap, Plug, and Adapter: Polypropylene
- Column Frit: Polyethylene
| |
| MEP HyperCel |
Purple |
90 µm |
2 - 12 |
> 20 mg/mL1 |
| HEA HyperCel |
Black |
90 µm |
2 - 12 |
40 mg/mL2 |
| PPA HyperCel |
Yellow |
90 µm |
2 - 12 |
40 mg/mL2 |
| |
Column Geometry
- Column Volume: 1.04 mL
- Bed Height: 1.48 cm (0.58 in.)
- Bed Diameter: 0.94 cm (0.37 in.)
Device Dimensions
- Diameter: 1.6 cm (0.6 in.)
- Length (Without Plugs): 4.8 cm (1.9 in.)
Connections
- Inlet: Threaded female luer lock
- Outlet: Rotating male luer locking hub
Recommended Flow Rates
Back Pressure
- Maximum: 3 bar (300 kPa, 43.5 psi)
Storage
1DBC determined using 5 mg/mL IgG in PBS; flow rate 60 cm/hr.; column dimension = 1.1 cm ID x 7 cm column; residence time (Tr) = 5.68 min.
2DBC at 10% breakthrough; 5 mg/mL BSA in PBS; flow rate 50 cm/hr.; column dimension = 1.6 cm ID x 3.75 cm; (Tr) = 4.51 min.
3DBC determined using 5 mg/mL Triton
High Purity of Monoclonal IgG From Ascites Fluid Using
MEP HyperCel Sorbent
Column run at flow rate of 70 cm/h with 50 mM Tris-HCl, pH used for binding and 50 mM acetate, pH 4 used for elution. In the curve, arrows indicate water (a) and sodium caprylate (b) washes. Arrow c indicates start of elution buffer. SDS-PAGE analysis (reduced conditions): 1 = crude sample; 2 = purified; H = heavy chain; L = light chain.
Influence of pH and Ionic Strength on the Binding Capacity of
MEP HyperCel Sorbent
IgG capacities obtained at 10% breakthrough on MEP HyperCel resin vs. pH (A) and ionic strength (B) of the binding buffer. Experimental conditions: column = 1.1 cm (0.4 in.) ID X 9 cm (3.5 in.); sample = IgG (2 mg/mL); flow rate = 90 cm/hr.; Tr = 7.8 min.
Distinct Separation of Standard Protein Mixture Using AcroSep HEA Columns
Peak a - Lysozyme
Peak b - α-Chymotrypsinogen A
Peak c - α-Chymotrypsinogen B
Peak d - BSA
Flow rate: 0.2 mL/min for sample injection and 0.5 mL/min for other steps. Carbonate buffer, pH 10.0 with 150 mM salt used for binding. Elution using mixed phosphate and citrate buffers of desired pH.
Distinct Separation of Standard Protein Mixture Using AcroSep PPA Columns
Peak a - Lysozyme
Peak b - α-Chymotrypsinogen A
Peak c - α-Chymotrypsinogen B
Peak d - BSA
Sample injection occurred at 0.2 mL/min then increased to 0.5 mL/min for the remainder of the procedure. PBS, pH 7.2 used for binding of proteins. Elution carried out using citrate buffers of respective pH and conductance as shown.
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