Performance evaluation of non-porous versus porous ion-exchange packings in the separation of proteins by high-performance liquid chromatography

Duncan, J.Keith; Chen, Albert J.C.; Siebert, Christopher J.

doi:10.1016/s0021-9673(01)84985-2

Cited by 31 publications

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“…The first IEC column packings for proteins were based on derivatized dextrans and agaroses (28). 'These were eventually replaced by animated and sulfonated polystyrenes and by high-23, December 1, 1994 performance stationary phases based on silica that were capable of supporting the higher pumping pressure required for high-speed chromatography (23,24). The most recent trend in IEC packings has been the development of a new class of pellicular resins (29).…”

Section: Iecmentioning

confidence: 99%

“…For example, the hydrocarbon chains of SDS can interact hydrophobically with the stationary phase and cause the sulfonate end of the molecules to be positioned at the packing-solution interface, turning the stationary phase surface into a strong cation exchanger (23). Alternatively, once the surfactant forms a sheath around the protein, the hydrocarbon exterior of that sheath may interact hydrophobically with the stationary phase (24).…”

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Chromatographic analysis of therapeutic proteins

Compton¹,

Kreilgaard²

1994

Anal. Chem.

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Analysis of J V Therapeutic ProteinsRapid LC applications HPLC has become a principal tool for the rapid separation and characterization of many classes of pharmaceuticals, including those based on peptides and proteins.Most HPLC methods for biomolecules use an aqueous-based mobile phase; the stationary phase can vary, depending on the chromatographic mode. Aqueous solvents are used because, in general, nonaqueous solvents denature proteins by interfering with their uniquely folded structure.With the emergence of new proteinand peptide-based drugs, it has become necessary to develop rapid LC methods that can separate and characterize these substances or be used for preparative purposes without disrupting the unique three-dimensional structure upon which pharmacological activity depends. In general, these methods include, but are not limited to, size exclusion chromatography (SEC), ion-exchange chromatography (IEC), hydrophobic interaction chromatography (HIC), reversed-phase LC (RPLC), and affinity chromatography (AC).Each method has its strengths and weaknesses; the appropriate one is selected on the basis of the structure of the Development of new pharmaceuticals is increasingly expensive.Not only are the costs of R&D, laboratory testing, clinical trials, administration, and overhead escalating, but the maturing of the pharmaceutical industry itself is adding to the burden. Ten years ago, the rule of thumb was that development of a new drug cost about $200 million. Today that figure has doubled. A drug patent gives its owner exclusive rights to the invention for 17 years. Because it often takes as many as 12 years to bring a drug to market, it is crucial that product development proceed as efficiently as possible. Today the general rule for evaluating return on investment for pharmaceutical products is that for each day's delay in bringing the product to market, $1 million in sales is lost.Analysis and purification are essential parts of drug R&D, manufacturing, and quality assurance. Using rapid analytical methods can significantly cut develop-

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