Weak partitioning chromatography for anion exchange purification of monoclonal antibodies

Kelley, Brian; Tobler, Scott A.; Brown, Paul E.; Coffman, Jon; Godavarti, Ranga; Iskra, Timothy; Switzer, Mary; Vunnum, Suresh

doi:10.1002/bit.21923

Cited by 121 publications

(77 citation statements)

References 24 publications

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“…This can enable a two chromatography recovery process comprising Protein A and anion exchange for many antibodies. 51 As with flow-through chromatography, the process is run isocratically, but, in contrast to flowthrough mode, the conductivity and pH are chosen such that the binding of both the product and impurities are enhanced, attaining an antibody partition coefficient (Kp) between 0.1-20, and preferably between 1 and 3. This takes advantage of the fact that the impurities to be removed are more acidic than the product.…”

Section: Chromatographic Processesmentioning

confidence: 99%

Recovery and purification process development for monoclonal antibody production

Liu

Winter

et al. 2010

mAbs

434

399

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Section: Chromatographic Processesmentioning

confidence: 99%

Recovery and purification process development for monoclonal antibody production

Liu

Winter

et al. 2010

mAbs

434

399

View full text Add to dashboard Cite

“…A third chromatography mode, called weak partitioning, is defined by K p values generally ranging from 1 to 10 (Kelley et al, 2008b;Vunnum et al, 2006). In this mode, impurity removal is generally superior to flow-through modes, while yield is only modestly reduced.…”

Section: Design Of Cha Chromatography Processesmentioning

confidence: 99%

High‐throughput screening of chromatographic separations: III. Monoclonal antibodies on ceramic hydroxyapatite

Wensel

Kelley

Coffman

2008

Biotech & Bioengineering

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High-throughput screening (HTS) of chromatography resins for identifying optimal protein purification conditions is becoming an integral part of industrial process development. In this work, ceramic hydroxyapatite (cHA) chromatography of 15 humanized monoclonal antibodies (mAbs) was examined by HTS. MAb binding, as quantified by partition coefficient (K p ), was measured under 92 combinations of sodium chloride, phosphate, and pH. Binding varied inversely with these variables for all mAbs tested. However, the magnitudes of binding among mAbs under identical conditions varied significantly, showing a >1.5 log range in K p . Analysis of variance (ANOVA) techniques were used to describe the binding of each mAb as a function of the three screen variables. Linear models relating log K p to the pH, log[sodium chloride], and log[phosphate] fit the data for each antibody with 93-96% accuracy. From these models, characteristic charge values for the cation exchange and metal coordination components of the multi-modal mAb/cHA interaction varied twofold across the mAbs, reflecting inherent variability in the number of contacts between a particular mAb and the cHA surface. Furthermore, we reduced the number of test conditions required from 92 to 8 while maintaining an accurate representation of the full binding response surface. This eight-point modeling method accurately predicted the binding behavior of mAbs as well as mAb aggregates, a common impurity in crude mAb preparations. Using this eight-point modeling method, binding and selectivity information for mAb and aggregate can be obtained from less than two milligrams of protein, making the method attractive for early manufacturability assessments.

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“…The SEC step can then both remove aggregates and change the buffer conditions. The following AIEX step can then be run in weak partitioning mode [15] to remove host cell proteins (HCP) and leached Protein A.…”

Section: Downstream Processing Platformmentioning

confidence: 99%

Model-Based Comparison of Antibody Dimerization in Continuous and Batch-Wise Downstream Processing

2015

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Monoclonal antibodies are generally produced using a generic platform approach in which several chromatographic separations assure high purity of the product. Dimerization can occur during the fermentation stage and may occur also during the downstream processing. We present here simulations in which a traditional platform approach that consist of protein A capture, followed by cation-exchange and anion-exchange chromatography for polishing is compared to a continuous platform in which dimer removal and virus inactivation are carried out on a size-exclusion column. A dimerization model that takes pH, salt concentration and the concentration of antibodies into account is combined with chromatographic models, to be able to predicted both the separation and the degree to which dimers are formed. Purification of a feed composition that contained 1% by weight of dimer and a total antibody concentration of 1 g/L was modeled using both approaches, and the amount of antibodies in the continuous platform was at least 4 times lower than in the traditional platform. The total processing time was also lower, as the cation-exchange polish could be omitted.

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Weak partitioning chromatography for anion exchange purification of monoclonal antibodies

Cited by 121 publications

References 24 publications

Recovery and purification process development for monoclonal antibody production

Recovery and purification process development for monoclonal antibody production

High‐throughput screening of chromatographic separations: III. Monoclonal antibodies on ceramic hydroxyapatite

Model-Based Comparison of Antibody Dimerization in Continuous and Batch-Wise Downstream Processing

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