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[Isolation of Pseudomonas aeruginosa exotoxin A by affinity chromatography using specific antibodies]
The method for the isolation of specific antibodies to P. aeruginosa exotoxin A from rabbit antitoxic serum by means of affinity chromatography has been developed. The conditions permitting the isolation of immunoglobulins to exotoxin A, practically without losses, have been experimentally established. The method for the isolation of exotoxin A from P. aeruginosa culture fluid with the use of specific antibodies to exotoxin A, immobilized on Sepharose 4B, has been worked out. This method makes it possible to obtain 21 mg of exotoxin A per cycle in a column having a working capacity of 25 ml, the yield being 87% of exotoxin A from its initial content in acellular culture fluid. The preparation of exotoxin A thus obtained is toxic, homogeneous and has specific enzymatic activity. Its molecular weight is 71 KD.
The method for the isolation of specific antibodies to P. aeruginosa exotoxin A from rabbit antitoxic serum by means of affinity chromatography has been developed. The conditions permitting the isolation of immunoglobulins to exotoxin A, practically without losses, have been experimentally established. The method for the isolation of exotoxin A from P. aeruginosa culture fluid with the use of specific antibodies to exotoxin A, immobilized on Sepharose 4B, has been worked out. This method makes it possible to obtain 21 mg of exotoxin A per cycle in a column having a working capacity of 25 ml, the yield being 87% of exotoxin A from its initial content in acellular culture fluid. The preparation of exotoxin A thus obtained is toxic, homogeneous and has specific enzymatic activity. Its molecular weight is 71 KD.