June 3, 1918 



Destruction of Tetanus Antitoxin 



493 



presence of the proteoses, peptones, etc., which make up the bulk of the 

 trypsin and pepsin preparations used, as explained on page 481. 



Mixture d.— The destruction of antitoxin by pepsin-hydrochloric 

 acid in this mLxture was obviously due to the hydrochloric acid, as the 

 results for mixture B indicate. Furthermore, these indicate that the pro- 

 teolysis in mixture D can not with certainty be regarded as the imme- 

 diate cause of the anti- 





^w 



20 



/ 2 3 ^ 

 n/9ys /p/GTsrsz? 



toxin destruction. 



Figure 4 shows the 

 simultaneous antitoxin 

 destruction and pro- 

 tein spHtting in mix- 

 ture D, experiment 22. 



Carri^re's (4) finding 

 that pepsin - hydro- 

 chloric acid does not 

 affect tetanus anti- 

 toxin is probably in- 

 correct. 



Mixture; e. — This 

 showed that the pep- 

 sin - hydrochloric acid 

 did not destroy ap- 

 preciable amounts of 

 the tetanus toxin. 

 The highest final con- 

 centration of hydro- 

 chloric acid in the toxin-antitoxin mixture injected was 0.005 P^r 

 cent in experiment 22. In experiments 20 and 21 it was 0.0027 P^r cent. 

 In these three experiments the dose of mixture E injected into one 

 guinea pig contained but 2 MLD. The deaths of the guinea pigs resulted 

 in the following number of hours after the subcutaneous injection of 

 the mixture: 66, 66, 66, 72, 74, 90. 



DESTRUCTION OF ANTHRAX IMMUNE BODIES BY PROTEOLYTIC 



ENZYMS 



Before the writers began the work on tetanus serum, they were en- 

 gaged in a similar study of anthrax serum. Inoculation experiments 

 were made for the purpose of ascertaining whether the immune bodies 

 in anthrax serum would be destroyed when the serum proteins were 

 digested by pepsin-hydrochloric acid or trypsin-sodium-carbonate. 

 The general method of the inoculation of the guinea pigs, the standardi- 

 zation of the virus, etc. have been described in a previous publication 

 (6, p. 41). Chemical analyses were made, similar to those mentioned 

 in the foregoing pages, for the purpose of measuring the extent of pro- 



FiG. 4. — The destruction of tetanus antitoxin ( X ) by pepsin- 

 hydrochloric acid; the digestion of coagulable protein past the coag- 



ulable stage ( Q ), and the Hberation of free amino nitrogen 



( 1 ). Mixture D, experiment 22. 



