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KMID : 0377619620030060591
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Korean Jungang Medical Journal
1962 Volume.3 No. 6 p.591 ~ p.597
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Effect of Heat on Hemagglutinin
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Abstract
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Effect of physical and chemical agents on antibody cativity has been studied by many worker as a basic problem in serology. Particularly as to effect of heat, many investingations were made by earlier workers, since this was a clue to solving the existiing problems related to preservation and purification of antiboddy and other fundamental facts in the serological field. Also it is a generally admitted fact that antibody is comparatively stable at low temperature, and with increasing temperature its activity gradually decreases.
The present study, howeve, deals with some curious phenomena newly observed which are not in accordance with the previous reports.
Rabbits weighing 2-2.5kg were divided into two groups, the 1st group was immunized against goat red cells, the 2nd group against chicken red cells. Ten ml of a 3% red cell suspension was injected intravenously eight times at four day intervals. Four days after each injection, immediately before the next injection, approximately 2 ml of blood was withdrawn for antibody titration.
the sera were diluted to 10 times with 0.85% sodium chloride solution(saline) or 0.5% phenolized physiological saline (phenilized saline) to prevent coagulation, placed in ampules, and heated at various tempeartures in a water bath. Antibody titer was determined by hemagglutination test.
The immune sera were heated at 56℃ to 84℃, at 2℃ intervals, for 30 minutes respectively, and hemagglutination titers were compared. While some of the antisera decreased in hemagglutinating activity with increasing temperature, as were reported by earlier workers, the other antisera showed the maximum titer on heating at 76℃ and 72℃ when the sera had been diluited with saline and phenilized saline respectively before heating, the titers being significantly higher than was inactivated at 56℃.
The sera were eated at 60˚, 64˚, 68˚,72˚ and 76℃ for 30 minutes, 1,2,4, 8, and 12 hours respectively and an unusual fact also was observed in some cases that prolinged heating made hemegglutination titers higher. For the sake of convenience such a urous phenomenon was erfered to as heat-activated hemagglutination, the serum as heat-activated hemagglutinating serum.
During the immunization course heat- activated hemagglutinating antibdy usually was not present in the earlier stage, but appeared in the later stage as immunizstion went on. This indicated that the appearance of heat- activated hemaggltinating antibody in immune serum had a close relationship with the progress of immunization.
The hemagglutination titers of immune sera heated at 72℃ or 76℃ had no relation to those heated at 56℃ and took an independent course during the whole immunization period, usually appearing later. There was difference in agglutination titers between the sera diluted with physiological saline and with phenolized saline prior to heating provided that heating the sera was limited within 30 minutes at less than 66℃. But higher temperatures or prolonged heating might cause a remarkable difference.
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