Formation of Antibodies in Fertile Hens Eggs

Abstract

The theoretical and practical interest inherent in the question of the time and location of antibody formation in the developing organism is responsible for a series of experiments in which fertile hens' eggs were repeatedly injected with foreign sera. After incubation for a certain time, the eggs were opened and precipitin tests were made on the remaining egg white. These tests were positive in approximately 4% of the cases. 20

The handling of the eggs during the process is quite delicate, and in spite of careful manipulation, a number of eggs were lost. Because of the obvious necessity of insuring a well-growing chick, the best grade of hatching eggs must be procured. The first injection is usually made on the third day of incubation, when fertility may be ascertained. The egg shell was first washed with mercuric bichloride and then with alcohol. Into each egg, from o.1 to 0.25 cc. of sheep serum [foreign soluble protein] which had previously been passed through a Berkefeld filter, was injected. In piercing the shell, care must be taken to cause as little disturbance of the contents as possible. The same precaution must be observed while forcing the foreign protein out of the syringe. After the injection had been made, the needle was retracted, and the hole immediately sealed with fairly thick collodion. These injections were repeated 3 or 4 times at intervals of 2 to 3 days, and the eggs were continuously incubated at 37.5°C, and handled as they would be for ordinary hatching purposes. After a total of 14 days' incubation, the eggs were opened and the egg white removed. It was diluted with 5 volumes of physiological salt solution, and filtered until perfectly clear.