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J Clin Microbiol. 1976 April; 3(4): 402-405

Elimination of bacteriophages from tissue culture serum by affinity chromatography.

ABSTRACT

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.