Factors influencing chromatography of proteins on short alkylsilane-bonded large pore-size silicas

M. J. O'Hare, M. W. Capp, E. C. Nice, N. H.C. Cooke, B. G. Archer

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The separation of proteins by high-performance gradient chromatography on short alkylchain bonded silica was studied with respect to pore size, mobile-phase composition, and temperature. Selectivity could be increased in particular cases by varying temperature or eluate compositon. Recovery of late-eluting, hydrophobic proteins was found to increase with flow rate and gradient slope. Recovery was also shown to be dependent on eluate composition-decreasing with added salt. The applicability of reverse-phase chromatography to proteins as large as 150 kD was demonstrated by the separation of monoclonal immunoglobulin.

Original languageEnglish
Pages (from-to)17-28
Number of pages12
JournalAnalytical Biochemistry
Issue number1
Publication statusPublished - 1 Jan 1982
Externally publishedYes

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