Improved immunoadsorption procedure with anion-exchange bacterial cell columns.
Bacterial cell columns for immunoadsorption were prepared with Streptococcus cells and triethylaminoethyl cellulose (Cellex-T) matrix material as a model system. Good column flow properties and satisfactory retention of the cells were obtained with ratios as high as 2 ml of packed cells/3 g dry weight of cellulose. Anion-exchange fractionation of whole serum by the Cellex-T was prevented by using 0.25 M NaCl in the developing buffer. Antibodies were adsorbed directly from whole serum and recovered in high yield by desorption at pH 2.3. Pre-exposing bacterial cells to formalin and washing them with acetone was necessary to ensure that they remained on the columns. One strain of Streptococcus salivarius (SS 908) was satisfactorily retained on a column only after cells were labeled with fluorescein isothiocyanate and washed with acetone. The means by which Cellex-T retains bacterial cells appears to be a combination of electronic attraction and physical entrapment.
D002852:Chromatography, Ion Exchange / Q000379:methods; D005455:Fluorescent Antibody Technique; D007163:Immunosorbent Techniques; D012965:Sodium Chloride / Q000494:pharmacology; D013291:Streptococcus / Q000276:immunology*
D012965:Sodium Chloride