B. Bednar et Jp. Hennessey, MOLECULAR-SIZE ANALYSIS OF CAPSULAR POLYSACCHARIDE PREPARATIONS FROM STREPTOCOCCUS-PNEUMONIAE, Carbohydrate research, 243(1), 1993, pp. 115-130
Purified capsular polysaccharide preparations from Streptococcus pneum
oniae serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F were analyzed by high
performance size exclusion chromatography (HPSEC) with multi-angle la
ser light scattering (MALLS), specific viscosity (SV), and refractive
index (RI) detection to determine the molecular size and molar mass of
each of the pneumococcal (Pn) polysaccharides. The M(w)'s of the poly
saccharides ranged from a low of 606 kg/mol for Pn4 to a high of 1145
kg/mol for Pn9V, and the z-average radii Of gyration ranged from 59 nm
for Pn14 to 72 nm for Pn18C. Estimations of molar mass of the highly
anionic polysaccharides (all but Pn14) by the universal calibration ap
proach were unsuccessful, resulting in a 27-53% overestimate of the M(
w)'s, though application of Mark-Houwink-Sakurada coefficients calcula
ted from the HPSEC-MALLS/SV/RI data resulted in estimates of M(w) that
were in agreement with the MALLS estimates for all but the Pn4 prepar
ation. These results emphasize the need for direct measurement of both
molecular size and intrinsic viscosity distributions for definitive c
haracterization of the molar mass, hydrodynamic volume, rigidity, and
drainage of complex biological polymers such as the pneumococcal polys
accharides.