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In Vivo Purification of I^sp131 Labeled Localizing Antirat Lymphosarcoma Antibody¹

From the Department of Radiation Biology, The University of Rochester School of Medicine and Dentistry, Rochester, New York

Abstract

Antibody preparations were separated from the sera of rabbits immunized with a homogenate of rat Murphy lymphosarcoma plus Freund adjuvant. When labeled with I¹³¹ and injected intravenously into rats bearing this tumor, they showed substantial tumor localization. One day after intravenous injection the portion of the injected I¹³¹ found in the tumors of 20 rats averaged 6.8% of the injected dose in an amount of tumor equal to 1% of the animals' weight. This I¹³¹ concentration in tumor was more than twice the blood concentration at time of sacrifice and several times the concentration found in any other perfused organ or tissue. The tumor localizing substance behaved like -globulin with starch electrophoresis. Control studies with labeled normal -globulins indicated that most of the accumulation of I¹³¹ in tumor was specific and due to antibody to the Murphy tumor.

The antibody purification procedure involved a step in which a labeled crude antibody preparation was injected into tumor bearing animals, the animals sacrificed a day later, and the I¹³¹ labeled antibody attached to the washed homogenate of these tumors eluted into solution with 32% urea in pH 8 borate buffer.

Preparations made by completely in vitro methods gave less specific tumor localization.

Footnotes

¹ This paper is based on work performed under contract with the U. S. Atomic Energy Commission at the University of Rochester Atomic Energy Project, Rochester, New York.