Purification of Type IV and Type VI Lipoteichoic Acids from Gram-Positive Mucosal Pathogens via a Two-Step Hot...

Type IV and Type VI lipoteichoic acids (LTAs) are found on the surface of Gram-positive mucosal pathogens. There is interest to purify, characterize, and evaluate these compounds as vaccine candidates. In traditional hot water-phenol extractions, Type IV and VI LTAs are found almost exclusively in the phenol fraction, and, due to the lengthy removal process, prolonged exposure to phenol can degrade them. Here we describe an efficient and time-saving new method to rapidly separate these LTAs from phenol through the addition of triethylammonium acetate (TEAA), and we present a continuous flow dialysis arrangement to rapidly remove excess phenol. We demonstrate the efficacy of this protocol through silver staining and immunoblotting of Type IV LTA from Streptococcus pneumoniae and Type VI LTA from Clostridium perfringens.