Exmere

    Exsil GFC family · 400 Å

    Exsil GFC 4000

    High performance size separation of high molecular weight proteins.

    400 Å5 µm60 K – 500 KpH 2 – 7.57000 psi
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    Core positioning

    Higher molecular weight range, same robust chemistry.

    Exsil GFC 4000 has been developed for high efficiency separation of higher molecular weight proteins with superior bed stability and economical performance.

    Technical specifications

    Product specifications.

    PhaseStable High Polarity
    Working pH rangepH 2 – 7.5
    Base silicaExsil 4000S, high purity silica
    Particle size5 µm
    Pore size400 Å
    Pore volume1.2 cc/g
    Cat. no.4000GU — Exsil GFC 4000, 5 µm
    Cat. no. (base silica)4000GSU — Exsil 4000S, 5 µm silica

    Key benefits

    Why Exsil GFC 4000.

    High stability

    Can be packed at 7000 psi.

    High efficiency

    Up to 100,000 plates/m for 5 µm / 8 mm ID.

    High recovery

    Recoveries in the 99 to 102% range.

    Wide applicability

    Low adsorption of basic proteins.

    Cost effective

    High performance at low cost.

    Reproducible

    Typical Exmere reliability.

    1 · Size exclusion range

    Shifted to higher molecular weight proteins.

    GFC of proteins on Exsil GFC 4000 shifts the retention of proteins to higher Kd values compared to Exsil GFC 3000. The Kd for albumin is in the region of 0.68, which compares well with the reported figure for TSK 4000SW of 0.70.

    The optimum operating range of Exsil GFC 4000 is 60 K – 500 K — significantly higher than the 15 K – 60 K range of Exsil GFC 3000. Together, the two columns cover the important 15 K – 500 K region.

    2 · Protein separations

    Excellent resolution in 8 mm ID and 4.6 mm ID formats.

    Thyroglobulin · Ovalbumin · Phenylalanine

    Exsil GFC 4000 — 8 mm ID (A) and 4.6 mm ID (B)

    Chromatogram of Thyroglobulin, Ovalbumin and Phenylalanine on Exsil GFC 4000 in 8 mm and 4.6 mm ID formats.

    A) 300 × 8 mm Exsil GFC 4000 at 1 mL/min and B) 250 × 4.6 mm Exsil GFC 4000 at 0.33 mL/min. Eluent: 50 mM potassium phosphate pH 7 / 150 mM sodium chloride.

    Reference chromatogram — Exmere technical note.

    3 · High speed & high resolution

    Mechanical headroom for faster or higher-resolution runs.

    Exsil GFC 4000 retains the excellent mechanical stability of Exsil GFC 3000 and can be packed at 7000 psi and operated up to at least 3000 psi. This opens up the opportunity to carry out either high speed or high resolution separations.

    The chromatograms opposite show a high speed separation completed on a 250 mm column in 3 minutes and a high resolution separation using three 250 mm columns. In both cases the back pressure was in the region of 2000 psi — well within the operating capabilities of Exsil GFC 4000, but around twice the maximum limit of some competitive materials.

    Thyroglobulin · Ovalbumin · Phenylalanine

    250 mm column (A) vs 750 mm column stack (B)

    High speed vs high resolution Exsil GFC 4000 separations.

    A) 250 × 4.6 mm Exsil GFC 4000 at 1 mL/min and B) 750 × 4.6 mm Exsil GFC 4000 at 0.33 mL/min. Eluent: 50 mM potassium phosphate pH 7 / 150 mM sodium chloride.

    Reference chromatogram — Exmere technical note.

    4 · Protein recovery & phase clean-up

    Robust to routine use and high-pH cleaning.

    Protein recovery

    Recovery was assessed using the acidic protein Human Serum Albumin and the basic protein Equine Cytochrome C using 50 mM potassium phosphate pH 7 / 150 mM sodium chloride eluent. In neither case was there evidence of protein loss, with recoveries in the 99 to 102% range.

    Phase clean-up

    Cleaning the column with 50 column volumes of 50% MeOH / 50% 10 mM NaOH had minimal effect on column performance. Restricted to 5 column volumes, the column could be cleaned a minimum of 10 times.

    Before / after high-pH clean-up

    Thyroglobulin · h-Serum Albumin · Phenylalanine

    Chromatogram before and after 50 column volumes of NaOH/MeOH clean-up on Exsil GFC 4000.

    A) New 250 × 4.6 mm Exsil GFC 4000 column and B) after 50 column volumes of 50% MeOH / 50% 10 mM NaOH. Eluent: 50 mM potassium phosphate pH 7 / 150 mM sodium chloride at 1 mL/min.

    Reference chromatogram — Exmere technical note.

    Not sure which Exsil GFC phase you need?

    Compare the other members of the Exsil GFC family.

    Exsil GFC 2000

    Coming soon

    Smaller-pore GFC phase for the lower MW segment.

    Pore size
    —
    Particle size
    5 µm
    Application range
    Coming soon
    Get notified

    Exsil GFC 3000

    Available

    For protein size separation in the lower to medium MW range.

    Pore size
    235 Å
    Particle size
    5 µm
    Application range
    15 K – 60 K
    View GFC 3000

    Exsil GFC 5000

    Available

    For very high molecular weight proteins.

    Pore size
    800 Å
    Particle size
    5 µm
    Application range
    150 K – 1250 K
    View GFC 5000

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