Abstract: | By combining gel permeation chromatography (GPC) and light-scattering spectroscopy, including photon correlation and angular distribution of absolute scattered intensity, we were able to characterize immunologically active Haemophilus influenzae type b polysaccharide (HIB Ps) bovine serum albumin (BSA) conjugates in terms of equivalent hydrodynamic radius rh ~ (6.2 ± 0.6) × 102 Å, apparent radius of gyration rg ~ (5.4 ± 0.3) × 102 Å, apparent molecular weight Mw ~ (3.5 ± 0.4) × 106 g/mol, and a second virial coefficient A2 ~ (1.9 ± 0.3) × 10?4 cm3 mol/g2. We could study the effects of each of the processes in the conjugate formation according to the following procedure: BSA (dialysis, modification, fractionation) + HIB Ps → HIB Ps/BSA conjugate (conjugate formation, fractionation). Narrow distributions of HIB Ps BSA conjugate formation can be achieved using fractionated BSA. |