OISHI, N.Y.; UEDA, C.M.P.M.; DIAS, W. de O.; LATRILHA, C.M.L.; RAW. I. The possible role of 
para-pepsin in the preparation of anti-venom sera. Mem. Inst. Butantan, 49 (21:41-45, 1987. 
Table 4 shows the effect of pepsin fractions, obtained by DEAE- 
cellulose chromatography of batch separation, on the anti-venom yield. It 
is clear that the caseinolytic activity at pH 3.2 is a better predictor of anti- 
venom yield activity, than that at pH 2.3 or 1.5. This suggests that the very 
large amount of crude pepsin which is used in anti-toxin preparation is due 
to the requirement of enough para-pepsin II, which is a minor component 
in commercial preparations. Spleen cathepsin D or calf stomach renin will 
not replace pepsin. 
Present results show that a partial purification of commercial pepsins 
would allow for the use of a much smaller concentration of enzyme in the 
preparation of anti-venom sera. This is important, as commercial pepsin, 
being crude stomach extracts may be contaminated with endotoxins. Fur- 
thermore, we have carried out a 5g/L crude pepsin throughout the process 
of serum purification, which results in a protein solution of about 1.2 g/L, 
still containing a detectable trace of pepsin activity. 
TABLE1 
EFFECT OF PEPSIN CONCENTRATION AND DIGESTION TIME ON SERUM YIELD 
pepsin 1 /20.000 
time 
pepsin 
total activity 
specific activity 
recovery 
min 
g/i 
mg venon/L serum 
activity/mg protein 
% 
— 
0 
400 
3.20 
100 
30 
0.001 
84 
15.84 
21 
30 
0.01 
104 
7.87 
26 
30 
0.1 
96 
8.00 
24 
30 
i 
80 
5.50 
20 
30 
5 
168 
7.53 
42 
90 
0.001 
2 
6.58 
15 
90 
0.01 
3 
3.27 
8 
90 
0.1 
4 
4.80 
12 
90 
1 
83 
5.10 
13 
90 
5 
65 
5.50 
14 
180 
0.001 
3 
7.10 
7 
180 
0.01 
4 
3.10 
10 
180 
1 
9 
5.80 
23 
180 
5 
13 
9.60 
32 
TABLE 2 
EFFECT OF PEPSIN CONCENTRATION ON COMPLEMENT FIXATION 
Pepsin 
1/20.000 g/L 
plasma dilution 
1/2 
1/4 
1/8 
1/16 
1/32 
1/64 
1/128 
0.5 
— 
— 
— 
_ 
_ 
_ 
_ 
0.25 
— 
— 
— 
— 
— 
— 
— 
0.10 
— 
— 
— 
— 
— 
— 
— 
0.05 
— 
— 
— 
— 
— . 
— 
— 
0.01 
— 
— 
— 
— 
— 
— 
— 
0.005 
— 
— 
— 
— 
— 
— 
— 
0.0025 
— 
— 
— 
— 
— 
— 
— 
0.00125 
4- + 
4 4 
+ + 
+ + 
— 
— 
— 
0.000625 
+ + 
4 + 
4 4 
4 4 
— 
— 
— 
0 
+ + + + 
4 4 4 4 
4 4 4 4 
+ + + + 
+ + + 
4 4 
- 
43 
cm 
SciELO 
11 12 13 14 15 16 17 
