Fig. 1
From: Hyaluronan is a natural and effective immunological adjuvant for protein-based vaccines
Synthesis and purification of HA-OVA. a Reaction scheme of deprotection of HA-acetal to HA-aldehyde (Step 1) and subsequent conjugation to a protein by reductive amination (Step 2); b 1H-NMR of HA200kDa-acetal in D2O. The modification degree of the polymer was calculated by comparing the integration value of the acetal moiety (α: 1.10 ppm) with that of the acetyl group of the HA backbone (β: 1.85 ppm); for correspondence of α and β, see the HA-acetal chemical formula in (a); c SEC-HPLC chromatograms of the HA-OVA conjugation reaction time course at 0 h and after 18 h. d HA-OVA after purification from the unreacted protein. The chromatographic profiles were obtained by SEC-HPLC on an analytical Zorbax GF-250 column (250 × 4.6 mm), eluted with 20 mΜ Na2HPO4 and 130 mM NaCl (pH 7.2) containing 20% (v/v) acetonitrile (ACN) at a flow rate of 0.3 mL/min. The eluate was monitored by measuring the absorbance at 280 nm
