Étude de l'élution non-linéaire en chromatographie en phase liquide préparative

Gareil, Pierre; Personnaz, L.; Feraud, Jean-Pierre; Caude, M.

doi:10.1016/s0021-9673(00)81841-5

Cited by 29 publications

(4 citation statements)

References 24 publications

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“…The major benefit of a FT operation compared to B/E is that the resin capacity is used to bind only the impurities and not the product, which is the major constituent of the load. Consequently, the relatively large process volumes may be applied with the ease and simplicity of an isocratic, volume overloaded operation (Gareil et al, 1980;Knox and Pyper, 1986).…”

Section: Introductionmentioning

confidence: 99%

Weak partitioning chromatography for anion exchange purification of monoclonal antibodies

Kelley

Tobler

Brown

et al. 2008

Biotech & Bioengineering

130

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Weak partitioning chromatography (WPC) is an isocratic chromatographic protein separation method performed under mobile phase conditions where a significant amount of the product protein binds to the resin, well in excess of typical flowthrough operations. The more stringent load and wash conditions lead to improved removal of more tightly binding impurities, although at the cost of a reduction in step yield. The step yield can be restored by extending the column load and incorporating a short wash at the end of the load stage. The use of WPC with anion exchange resins enables a two-column cGMP purification platform to be used for many different mAbs. The operating window for WPC can be easily established using high throughput batch-binding screens. Under conditions that favor very strong product binding, competitive effects from product binding can give rise to a reduction in column loading capacity. Robust performance of WPC anion exchange chromatography has been demonstrated in multiple cGMP mAb purification processes. Excellent clearance of host cell proteins, leached Protein A, DNA, high molecular weight species, and model virus has been achieved.

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Section: Introductionmentioning

confidence: 99%

Weak partitioning chromatography for anion exchange purification of monoclonal antibodies

Kelley

Tobler

Brown

et al. 2008

Biotech & Bioengineering

130

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“…There are two forms of overload that are generally experienced in LC systems; mass and volume. Fundamental considerations on mass and volume overloading have been discussed in detail and some general consensus has long existed . As a signature response under mass overloaded conditions, the front of an eluting peak becomes sharp while pronounced tailing is exhibited.…”

Section: Resultsmentioning

confidence: 99%

“…Fundamental considerations on mass and volume overloading have been discussed in detail and some general consensus has long existed. 8,[57][58][59][60][61] As a signature response under mass overloaded conditions, the front of an eluting peak becomes sharp while pronounced tailing is exhibited. In addition, the breakthrough time of the peak front is shortened; this is regarded as a "fronting" situation.…”

Section: Mass and Volume Overloading In Protein Separationsmentioning

confidence: 99%

Overload Effects in Reversed Phase Protein Separations using Capillary‐Channeled Polymer Fiber Columns

Wang

Marcus

2018

Biotechnology Progress

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In preparative chromatography, overloading effects are a fact of life. However, peak distortion is problematic in analytical scale chromatography, especially in the case of bio-macromolecule separations. Capillary-channeled polymer fibers have been employed for fast protein separations in reversed phase, ion exchange, and hydrophobic interaction liquid chromatography. Although the primary advantage of the phase is operation at high linear velocities (~100 mm s ) without van Deemter C-term limitations, the limited specific surface area (<5 m g ) suggests that a thorough understanding of overloading effects is needed. We evaluate important factors (injected mass and volume) affecting overload in terms of peak height, width and shape (asymmetry). Overload conditions readily apparent as the peak shape changes from a Gaussian distribution to a left-triangle with slight tailing; more-or-less classical overload behavior. Three methods were used to compute column efficiency in terms of plate counts (N), including the area height, half-height, and the Dorsey-Foley approaches. The half-height method is best-suited for describing column overload, accounting for peak distortion and achieving high levels of consistency. The limiting plate count (N ) and the column sample loading capacity (ω ) were key metrics to characterize overall column performance. Peak capacity (P) was used to assess gradient elution separation performance. Increased flow rates and column length result in enhanced loading capacity. pH mismatch between the sample matrix and the mobile phase, a common cause of protein overload with other phases, was not responsible for distorted peak shapes unless the solvent pH is close to the pI of protein samples.

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“…

Band broadening and deviations from symmetry which are generally observed when a chromatographic column is overloaded are mainly caused by the nonlinearity of the solute-stationary-phase equilibrium isotherm. For moderate concentrations (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) µ /cm1 23) the isotherm can most often be replaced by a two-term expansion. Then the system of mass balance equations of chromatography can be solved and an analytical equation of the peak profile obtained.

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mentioning

confidence: 99%