Elimination of bacteriophages from tissue culture serum by affinity chromatography.
Use of immunoadsorbents to remove bacteriophages from tissue culture serum was investigated. Immune globulins from rabbit antiserum prepared against phi V-1 phage were immobilized by covalent linkage to activated porous silica glass derivatives of p-aminoarylamine and to Sepharose-4B. Chromatographic columns of each material were used to filter samples of a fetal bovine serum into which had been introduced 8100 plaque-forming units of the phage per ml. Efficiency of removal was determined by plaque assays of phi V-1 phage recovered in the effluent fluids. Activated but uncoupled matrices nonspecifically removed from 49 to 59% of the phages introduced into the experimental serum. A reduction of 35 to 37% in phage content occurred in the serum after filtration through columns coupled to nonantibody protein. With specific immune globulins attached, the Sepharose-4B matrix reduced the concentration of phage in the serum below a detectable quantity. Noapparent alterations occurred in the growth-promoting property of serum filtered through the Sepharose-4B immunoadsorbent as measured by cloning efficiency of BHK-21, WI-38, and FRhL-2 cells. These experiments serve as a model system for use of immunoadsorbents for selective removal of bacteriophages and perhaps other extraneous microbial agents from tissue culture serum.
D000327:Adsorption; D000818:Animals; D000914:Antibodies, Viral; D001435:Bacteriophages / Q000302:isolation & purification*; D002417:Cattle; D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods; D003470:Culture Media; D005312:Fetal Blood; D007136:Immunoglobulins; D011817:Rabbits / Q000276:immunology; D012685:Sepharose
D000914:Antibodies, Viral; D003470:Culture Media; D007136:Immunoglobulins; D012685:Sepharose