

1905. | Chemistry of the Toxin-Antitoxin Reaction. 187 
latter has a fairly sharp boundary against the unhemolysed and turbid 
suspension. The experiments were carried out at about 18° C. The 
gelatine suspensions were contained in tubes of 1 cm. diameter, and were 
10 cm. in length. The test volume of supernatant fluid was 1 c.c. 
Lysin.—The rate at which hemolysis proceeded from the surface decreased 
when the concentration of the lysin decreased from 1 to 0°0625 per cent., 
but lower concentrations down to 0:0078 per cent. gave almost the same effect 
as 0:0625 per cent. or low concentrations, then, the rate of the hemolytve effect 
as not dependent on the mass of the lysin. Controls with saline gave no trace 
of hzmolysis.* 
Antilysin.—A layer of the gelatine blood suspension 2 mm. thick was 
placed on a gelatine column containing 10 per cent. antilysin, and after 
standing for 40 hours at 18° C., a gelatine layer containing 0°5 per cent. lysin 
was superimposed. The blood was completely hzemolysed in 40 hours, 
whereas in a control with gelatine saline no hemolysis took place. When, 
however, antilysin was mixed with the gelatine suspension, so that the 
concentration was 2°5 per cent., it was found that a gelatine layer containing 
0-5 per cent. lysin failed to give a trace of hemolysis in 40 hours. It follows, 
therefore, that the antilysin in the first experiment could not have been 
present in a concentration of 2°5 per cent. even in the layer immediately in 
contact with the 10-per-cent. antilysin in gelatine. 
Again, as 0°5 per cent. lysin hemolyses 2 mm. in 40 hours under normal 
conditions, no retardation has taken place, and it is probable that megatheriwm 
antilysin does not diffuse appreciably through gelatine. 
Mirtures.—A neutral mixture, containing 0°5 per cent. precipitated lysin 
and 3°73 per cent. antilysin in saline gelatine (Table IV, No. 2), and an over- 
neutralised mixture, No. 1, containing 0°5 per cent. lysin and 5 per cent. anti- 
lysin, after being brought to equilibrium, showed the presence of at least a trace of 
free lysin. The hemolytic effects were equal and of the magnitude for which 
Table Il]. —Diffusion of Megatherium Lysin through Gelatine Columns, 
showing Hemolysis, in Millimetres. 









Percentage of f F : : i : ' Saline 
lysin ...... 1 0° | 0°25 | 0:125 | 0:0625 | 0:0817 | 0°0159 | 0:0078 imate 
GO Dys. 2.0.06 ero Ou) 1 eZ) Le 1:0 1:0 1:0 1:0 0:0 
oe 4°513°5| 2:2 |1°5 1:0 1:0 1°0 1:0 0:0 
* Compare Table III. 
