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Figure 4 | BMC Biotechnology

Figure 4

From: Improving the large scale purification of the HIV microbicide, griffithsin

Figure 4

Summary of the effects of 0.1 M MgCl 2 and 10 mg/mL bentonite on GRFT extract purity. a) Coomassie stained SDS-PAGE showing the initial extraction of GRFT (1), subsequent pH adjustment to pH4 (2) and heating to 55°C followed by centrifugation (3). The resulting supernatant was untreated (control) or treated with MgCl2 and/or bentonite and stirred overnight at 4°C. The following samples are the resulting supernatants; (4) Control −4°C, (5) 0.1 M MgCl2, (6) 10 mg/mL Bentonite, (7) 0.1 M MgCl2 & 10 mg/mL Bentonite . The following samples are the resulting pellets after treatment; (8) Pellet - 4°C, (9) Pellet - 0.1 M MgCl2, (10) Pellet - 10 mg/mL Bentonite, (11) Pellet - 0.1 M MgCl2 & 10 mg/mL Bentonite. b) Comparison of gp-120 binding GRFT concentrations relative to the control sample (4). Determined through a gp-120 binding ELISA and represented as a percentage of functional GRFT in the control extract. c) Comparison of the TMV coat protein concentration relative to the control extract. Determined through densitometry measures of TMV specific westerns and represented as a percentage of TMV in the control extract. b and c both analyzed by a one-way ANOVA with Bonferroni’s multiple comparison test comparing all groups. * represents a post-hoc test with a p-value < 0.05. Table: Densitometry readings of SDS-PAGE represented in a. The shaded area represents an extract condition achieving greater than 80% purity of GRFT.

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