Japanese Journal of Veterinary Research;Volume 2, Number 4

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BIOCHEMICAL STUDIES ON THE INCREASE AND FORMATION OF ANTIBODIES WITH CHYMOPAPAIN IN VITRO : I. STUDIES ON THE INCREASE OF DIPHTHERIAL ANTITOXIC TITERS FROM THE PEPSIN-DIGESTED ANTITOXIC GLOBULINS

ITO, Tokiya

Permalink : http://hdl.handle.net/2115/1662
JaLCDOI : 10.14943/jjvr.2.4.143

Abstract

Purified eu- and pseudoglobulin solutions of equine diphtherial antitoxic sera obtained at the initial stages of the immunization, were incubated with pepsin at 37℃, pH 4.4 and Eh 200〜250 mV. In 72〜108 hours, pH was adjusted to 7.3,and the solutions were incubated further mixed with purified oxidized chymopapain, purified concentrated diphtherial toxin being supplied hydrogen peroxide and other oxidizing agents at 40℃ and Eh 450〜500mV. In 48〜84 hours, when the protein-N values reached their highest, the mixtures were made up to 60% saturation of ammonium sulfate at pH 8.0. The formed precipitates were electrodialyzed and finally concentrated to the volumes equivalent to those of the corresponding starting native globulin solutions. The antitoxic titers of these preparations were found to be 4 to 7 times as great as those of the each starting native globulins by RAMON'S flocculation test and by protective test using guinea pigs. These preparations, prepared by these procedures, are consisted of largely small but complete avid and less antigenic antitoxins.

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