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Size-exclusion chromatography in an analytical perspective

1993; Elsevier BV; Volume: 648; Issue: 1 Linguagem: Inglês

10.1016/0021-9673(93)83284-y

1873-3778

Lars Hagel,

Protein purification and stability

Modern media for size-exclusion chromatography combine maximum selectivity with high plate count to resolve solutes differing in molecular mass by as little as 20%. Separations of solutes differing in molecular mass by only 10% will be very hard to accomplish even for rod-shaped molecules. The reduced column lengths and porosity of small-particle-sized media unfortunately reduces the inherent resolving power to the same order as traditional columns but with a substantial reduction in analysis time. The use of well-calibrated columns for obtaining estimates of molecular mass yields a precision close to that of absolute methods. The simplicity and accuracy of the integral calibration method using a sample of broad molecular mass distribution makes this procedure very suitable for manual calibration of size-exclusion columns.