Microspheres for Separation of Bioactive Small Molecules

Purification of Modified Therapeutic Proteins Available on the Market: An Analysis of Chromatography-Based Strategies

“…Studies have shown that the change in hydrophobicity between the native and PEGylated proteins is sufficient to separate these species ( Cisneros-Ruiz et al, 2009 ). However, the hydrophobicity difference that allows an efficient separation by HIC among mono- and multi-PEGylated species (including positional isomers) is only large enough when the attached PEG has a high molecular weight (>20 kDa) ( Mayolo-Deloisa et al, 2012 ; Huang et al, 2013 ). Hence, HIC may not exhibit a clearly defined elution profile of mono- and multi-PEGylated species or positional isomers when small PEG molecular weight is used ( Pfister and Morbidelli, 2014 ), holding back the use of HIC in large-scale purification of PEGylated proteins in comparison to IEX or SEC.…”

“…Features such as backbone matrix, ligand, particle size distribution, and pore size will determine the physical and chemical properties of the chromatographic media. To achieve a good performance in the purification of PEGylated proteins, the chromatographic resin should meet several aspects such as high hydrophilicity, good chemical and physical stability, large pore size according to the target PEGylated protein, high binding capacity and resolution, narrow particle size distribution, and avoid non-specific interactions ( Huang et al, 2013 ).…”

“… AEX, anion exchange chromatography; CEX, cation exchange chromatography; CLA, cross-linked agarose; CT, chromatographic technique; FF, Fast Flow; HIC, hydrophobic interaction chromatography; HP, high performance; NA, information not available; SEC, size exclusion chromatography. a Avallin et al, 2016 b Miyabe and Guiochon, 2000 c Mant and Hodges, 1991 d Yao and Lenhoff, 2006 e Yao and Lenhoff, 2004 f Huang et al, 2013 g Nweke et al, 2017 . …”

“…Acrylamide-dextran copolymer used in MacroCap SP resin, is another matrix worth mentioning. MacroCap SP was designed especially for the purification of PEGylated molecules at large-scale, with operating conditions at 120 cm/h, with an optimum bed height of 15–30 cm ( Fee and Van Alstine, 2011 ; Huang et al, 2013 ).…”

“…On the contrary, small particles (20–50 μm) are usually employed for large-scale purifications that require a high efficiency ( Carta and Jungbauer, 2010 ). Beads with a narrow particle size distribution (PSD) may result in less backpressure and higher resolution ( Huang et al, 2013 ). This is the case for resins such as Source 15Q, Mono Q and POROS 50 HQ, which were designed for fast high-resolution ion exchange protein separations.…”