Chemical, physical and chromatographic properties of Superdex 75 prep grade and Superdex 200 prep grade gel filtration media

Kägedal, Lennart; Engström, Björon; Elelgren, Hans; Lieber, Anna-Karin; Lundström, Håkan; Sköld, Annelie; Schenning, Mikael

doi:10.1016/s0021-9673(01)88883-x

Cited by 25 publications

(5 citation statements)

References 10 publications

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“…This is illustrated in Figure 9 which also shows the impact of the derivatisation of the base matrix for producing Superdex 200. The selectivity change is caused by covalently attaching dextran to the agarose skeleton (29). This will reduce the conformational entropy of the solutes entering the porous network and thus decrease the distribution coefficient, and as seen from Figure 9 the effect is more pronounced for solutes of increasing molecular size.…”

Section: Evaluation Of the Properties Of Novel Media For Size Exclusimentioning

confidence: 98%

“…The properties of a new family of media, designed for high resolution aqueous size exclusion chromatography of small and medium size biomolecules was evaluated with respect to theoretical attainable functional performance. The media, Superdex, is a composite material where dextran is covalently bound to the backbone of crosslinked agarose and the molecular mass and amount of dextran incorporated was adjusted to yield the desired selectivities (29). Data for this type of medium is given in Table I.…”

Section: Evaluation Of the Properties Of Novel Media For Size Exclusimentioning

confidence: 99%

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Characteristics of Modern Media for Aqueous Size Exclusion Chromatography

Hagel

1996

ACS Symposium Series

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Important characteristics of media for size exclusion chromatography include particle size, pore volume, sorptive properties and matrix rigidity. However, the single most decisive parameter is the optimization of the selectivity to cope with the demands of high resolution separations. Whereas traditionally media are manufactured to encompass a broad separation range we now see a focus to a few key applications. These include a renewed interest in separation of smaller macromolecules, e.g. peptides and protein fragments and in dedicated separations such as monoclonal antibodies from growth media additives. The increase in selectivity may be obtained by selective grafting of polymers to reduce accessibility of pores to a small size range of solutes and in principle mimic the functionality of a single pore size support. The properties of such a medium designed for aqueous size exclusion chromatography, Superdex, is reviewed to elucidate the effect of important characteristics.By definition, aqueous size exclusion chromatography is carried out using a porous support. The molecules are separated due to the varying degrees of steric exclusion from the pore volume as caused by different molecular sizes of the solutes. This will result in the molecules being eluted from the column in order of decreasing size. Furthermore, the separation will only take place over an elution volume corresponding to the pore volume. Thus, the effective separating volume in aqueous size exclusion is small as compared to adsorptive techniques (i.e. solutes will ideally only be eluted within one column volume). In preparative purifications this limitation is generally of little importance since the desired separation is achieved through employing a ge)l medium with a suitable selectivity. Also for analytical applications where one component is of major interest it is often possible to find a gel of matching selectivity. However, the analysis of broad molar mass distributions may demand a wide separation range to encompass the whole distribution and this may be obtained by

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Section: Evaluation Of the Properties Of Novel Media For Size Exclusimentioning

confidence: 98%

Section: Evaluation Of the Properties Of Novel Media For Size Exclusimentioning

confidence: 99%

Characteristics of Modern Media for Aqueous Size Exclusion Chromatography

Hagel

1996

ACS Symposium Series

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“…Using water as eluent, most of the detected compounds were eluted close to the exclusion limit (void limit). The cation exchange capacity of Superdex 75 HR is reported to be low (Kagedal et al, 1991;Hils et al, 1999). However, it seems to be large enough to cause electrostatic repulsion between the gel and the organic compounds of the sample.…”

Section: Resultsmentioning

confidence: 99%

The effect of two kinds of eluents on the speciation of Al, Mn, and Fe by size-exclusion chromatography of some Brazilian soil solutions

Pérez

Campos

2005

Chemical Speciation & Bioavailability

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Size-exclusion chromatography is a powerful tool for distinguishing dissolved organic matter in relation to its molecular size. However, problems may arise in the choice of the most appropriate eluent composition, regarding the gel used. In the present work, soil solution samples from four Brazilian topsoils were analyzed by high-performance liquid chromatography (HPLC) using a size-exclusion polymer-based column (Superdex 75 HR 10y30, Pharmacia). Water and 0.01 mol L À1 NaCl were investigated as mobile phases. The analysis of the UV peak distribution as well as their corresponding Fe, Mn, and Al concentrations suggested that the salt solution should be preferred since it minimizes unwanted interactions between the sample and the support material. Fe, Mn, and Al speciation calculated by the Geochem-PC program involving the Mattigod and Sposito Mixed Model was compared with the Fe, Mn, and Al-organic complex concentrations determined by SEC-HPLCyGFAAS. No correlation was found for Mn and Fe. For Al a significant non-linear correlation was observed.

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“…In case of Superdex 75 column, the area under elution profile up to 16 min represented aggregates, area under the peak from 16 to 19 min represented monomer and the area under the profile beyond 19 min showed the presence of fragments. Superdex 75 shows an optimal separation range from 3 kDa to 70 kDa [19][20][21]. The AUC corresponding to aggregates and fragments were integrated to obtain the percentage of aggregates and fragments in the supernatant fraction of the sample (SI Fig.…”

Section: Size-based Characterization Of Stressed Mab Samplesmentioning

confidence: 99%