Figures & data
Figure 1. Chromatogram of (a) B. asper venom, (b) venom adsorbed on aluminum hydroxide, (c) venom adsorbed on calcium phosphate, or (d) venom co-precipitated with calcium phosphate. Separation was performed by reverse-phase HPLC using a Lichrosphere RP 100 C18 column (250 × 4 mm, 5 µm particle size). Fractions were eluted at 1 ml/min with a linear gradient (discontinuous line) of water and acetonitrile as outlined in the Materials and methods. Detection was set at 215 nm. Fractions were numbered according to Alape-Girón et al. (Citation2008).
![Figure 1. Chromatogram of (a) B. asper venom, (b) venom adsorbed on aluminum hydroxide, (c) venom adsorbed on calcium phosphate, or (d) venom co-precipitated with calcium phosphate. Separation was performed by reverse-phase HPLC using a Lichrosphere RP 100 C18 column (250 × 4 mm, 5 µm particle size). Fractions were eluted at 1 ml/min with a linear gradient (discontinuous line) of water and acetonitrile as outlined in the Materials and methods. Detection was set at 215 nm. Fractions were numbered according to Alape-Girón et al. (Citation2008).](/cms/asset/b264cdb9-0ff4-41da-afb8-29dfe1f3b986/iimt_a_772267_f0001_b.jpg)
Table 1. Capacity of mineral salts to couple venom proteins.
Figure 2. Recruitment of leukocytes to the peritoneal cavity of mice injected with venom: dissolved in PBS (▾); adsorbed on aluminum hydroxide (•); adsorbed on calcium phosphate (▪); or co-precipitated with calcium phosphate (▴) (see Materials and methods for experimental details). Results are expressed as mean of quadruplicates (±SD). *Value significantly different from venom/PBS value (p < 0.05).
![Figure 2. Recruitment of leukocytes to the peritoneal cavity of mice injected with venom: dissolved in PBS (▾); adsorbed on aluminum hydroxide (•); adsorbed on calcium phosphate (▪); or co-precipitated with calcium phosphate (▴) (see Materials and methods for experimental details). Results are expressed as mean of quadruplicates (±SD). *Value significantly different from venom/PBS value (p < 0.05).](/cms/asset/8035cc2c-eb43-44ef-beda-dc2bc3e2f12e/iimt_a_772267_f0002_b.jpg)
Figure 3. Effects of adjuvants coupled to B. asper venom proteins on phagocytic activity of macrophages. Phagocytosis of zymosan particles was determined by phase-contrast microscopy (see Materials and methods for experimental details). Macrophages (harvested 96-h after intra-peritoneal injection of thioglycollate) were challenged with immunogen for 60 min before addition of zymosan. Bars correspond to experiments in which the venom was (a) dissolved in PBS, (b) co-precipitated with calcium phosphate, (c) adsorbed on calcium phosphate or (d) adsorbed on aluminum hydroxide. Values represent mean (±SE) from four mice. *Value significantly different from venom/PBS value (p < 0.05).
![Figure 3. Effects of adjuvants coupled to B. asper venom proteins on phagocytic activity of macrophages. Phagocytosis of zymosan particles was determined by phase-contrast microscopy (see Materials and methods for experimental details). Macrophages (harvested 96-h after intra-peritoneal injection of thioglycollate) were challenged with immunogen for 60 min before addition of zymosan. Bars correspond to experiments in which the venom was (a) dissolved in PBS, (b) co-precipitated with calcium phosphate, (c) adsorbed on calcium phosphate or (d) adsorbed on aluminum hydroxide. Values represent mean (±SE) from four mice. *Value significantly different from venom/PBS value (p < 0.05).](/cms/asset/226b2814-d247-4242-9238-7e2432c2af9a/iimt_a_772267_f0003_b.jpg)
Figure 4. Antibody response towards fractions (a) 9–11, (b) 17–19, (c) 22–23, and (d) 26–30 of B. asper venom, induced by venom dissolved in PBS (▾), adsorbed on aluminum hydroxide (•), adsorbed on calcium phosphate (▪), or co-precipitated with calcium phosphate (▴). *Value significantly different from venom/PBS value (p < 0.05).
![Figure 4. Antibody response towards fractions (a) 9–11, (b) 17–19, (c) 22–23, and (d) 26–30 of B. asper venom, induced by venom dissolved in PBS (▾), adsorbed on aluminum hydroxide (•), adsorbed on calcium phosphate (▪), or co-precipitated with calcium phosphate (▴). *Value significantly different from venom/PBS value (p < 0.05).](/cms/asset/c9772d4c-f540-4ea3-adfb-7f6221a18874/iimt_a_772267_f0004_b.jpg)