BMC Biotechnology最新文献

Background: Echis ocellatus is a highly venomous snake that can cause serious medical complications due to the presence of toxic proteins in its venom. These proteins, such as echicetin and phospholipase A₂ (PLA2), often cause severe pathophysiology in snakebite victims. Ganoderma lucidum is recognised for its pharmacological benefits against various diseases. However, the potential of this fungus as an antivenom has not yet been reported.

Objective: This study investigated the inhibitory effects of G. lucidum on haemorrhagic and anticoagulant activities induced by E. ocellatus venom, and identified its possible bioactive inhibitor compounds using in vitro, in vivo, and in silico methods.

Methods: Ganoderma lucidum was extracted using methanol in a standard procedure, and varying doses (20 and 40 mg) of the extract were tested against the biological activities E. ocellatus venom. Thereafter, the extract of the G. lucidum was subjected to Gas Chromatography-Mass Spectrometry (GC-MS) analysis to identify its bioactive compounds. The identified compounds were docked against the catalytic active sites of echicetin and PLA2 proteins to determine the best inhibitor compound. The Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) properties of compounds were determined using the ADMETlab 2.0 web tool.

Results: The extract caused 62.96 ± 1.03% and 59.25 ± 1.59% venom-induced haemorrhage inhibition at doses of 20 and 40 mg, respectively, while plasma clotting times were shortened to 132 and 163 s at 20 and 40 mg, respectively. The GC-MS identified 29 bioactive compounds from G. lucidum extract, out of which hesperidin had the highest docking scores of - 9.3 kcal/mol and - 9.9 kcal/mol against the catalytic sites of echicetin and PLA2 enzymes, respectively.

Conclusion: The results indicate that G. lucidum has antivenom potential against E. ocellatus venom-induced toxic activities, and identified hesperidin as a promising compound for antivenom exploration against viper envenoming.

Background: The World Health Organization (WHO) recommends combination therapy with two non-overlapping monoclonal antibodies (mAbs) to enhance protection in Rabies post-exposure prophylaxis (PEP). While bispecific antibodies (bsAbs) provide superior protection over monoclonal antibodies (mAbs) by simultaneously targeting two distinct epitopes, their development is hindered by complex and inefficient vector design. In this study, we aimed to develop a novel single-vector expression system to efficiently produce potent anti-rabies virus (RABV) neutralizing bsAbs.

Results: We engineered a minimalistic peaSKYA single-vector featuring bidirectional dual-CMV promoters to enable synchronous co-expression of antibody heavy (HC) and light chains (LC) within a single construct. By eliminating redundant backbone elements, this optimized vector increased the expression-cassette ratio by 17% and improves transfection efficiency by 32% compared to dual-vector systems. The platform demonstrated 2-fold greater yields of bispecific antibodies in HEK293F transient expression, with the tetravalent DVD-Ig format antibody HHDVD showing significantly enhanced neutralization potency versus parental monoclonal antibodies in both pseudovirus-based neutralization assays (PBNA, 2193.19 IU/mg) and rapid fluorescent focus inhibition tests (RFFIT, 1679.52 IU/mg).

Conclusions: We developed peaSKYA single-vector expression system for robust production of bispecific antibodies, with candidate HHDVD exhibiting potential in Rabies PEP. This streamlined vector design approach exhibited remarkable potential in bispecific neutralizing antibody development, offering a transferable platform for other engineered bispecific antibodies.

Patulin (PAT), a mycotoxin produced primarily by Penicillium expansum, poses significant health risks and frequently contaminates apples and apple-derived products, often exceeding permissible safety limits. This study investigated the potential of orotate phosphoribosyl transferase (URA5) to degrade PAT in apple juice under controlled conditions. PAT degradation was assessed at initial concentrations of 100 µg/L and 250 µg/L, with enzymatic treatment using 0.2 mg/mL URA5. Samples were incubated for up to 24 h, and PAT degradation was monitored at time intervals of 3, 6, 9, 12, 18, and 24 h using liquid chromatography-mass spectrometry (LC-MS). The results demonstrated a time-dependent PAT degradation, with significant reductions observed as incubation time increased. After 6 h, PAT concentrations decreased to 57.30 µg/L and 112.69 µg/L for the 100 µg/L and 250 µg/L samples, respectively. At 12 h, PAT levels in the 100 µg/L sample fell just below the permissible limit (50 µg/kg), while substantial degradation was observed in the 250 µg/L sample. By 18 h, PAT concentrations dropped further to 47.22 µg/L and 40.10 µg/L, reaching safe consumption levels. After 24 h, degradation rates reached 96.36% and 98.25%, reducing PAT levels to 30.22 µg/L and 31.48 µg/L, confirming the efficacy of URA5 in detoxifying PAT-contaminated apple juice. The findings highlight the potential application of URA5 as a biocatalyst for PAT detoxification in the fruit juice industry. Compared to existing detoxification methods, enzyme-based degradation presents a promising, environmentally friendly, cost-effective, and non-toxic alternative. Further studies should explore its feasibility in large-scale processing and its interaction with other contaminants in commercial apple juice production.