Efficient in planta production of amidated antimicrobial peptides that are active against drug-resistant ESKAPE pathogens.

Antimicrobial peptides (AMPs) are promising next-generation antibiotics that can be used to combat drug-resistant pathogens. However, the high cost involved in AMP synthesis and their short plasma half-life render their clinical translation a challenge. To address these shortcomings, we report efficient production of bioactive amidated AMPs by transient expression of glycine-extended AMPs in Nicotiana benthamiana line expressing the mammalian enzyme peptidylglycine α-amidating mono-oxygenase (PAM). Cationic AMPs accumulate to substantial levels in PAM transgenic plants compare to nontransgenic N. benthamiana. Moreover, AMPs purified from plants exhibit robust killing activity against six highly virulent and antibiotic resistant ESKAPE pathogens, prevent their biofilm formation, analogous to their synthetic counterparts and synergize with antibiotics. We also perform a base case techno-economic analysis of our platform, demonstrating the potential economic advantages and scalability for industrial use. Taken together, our experimental data and techno-economic analysis demonstrate the potential use of plant chassis for large-scale production of clinical-grade AMPs.

Detection of the administration of human erythropoietin (HuEPO) to canines.

Recombinant human erythropoietin (rHuEPO) doping is prohibited in animal (canine and equine) sport. The effectiveness of a range of immunoassay screening methods for the detection of rHuEPO in canine urine was evaluated. The excretion profiles following rHuEPO administration to dogs were investigated. The presence of rHuEPO in postadministration samples was confirmed using the World Anti-Doping Agency (WADA)-approved isoelectric focusing immunoblotting confirmatory technique. Following the administration study, a screening program involving approximately 6000 greyhound sport (mostly racing) samples was undertaken for rHuEPO. This resulted in the detection of the first rHuEPO positives in the world of canine or equine sport. In an additional case, endogenous HuEPO was detected in a sample submitted as greyhound urine. It was determined that this arose from the submission to control stewards, as greyhound urine, of a substance that was, in fact, human urine. This was a particularly welcome development as definitive confirmatory evidence of such sample switching can be difficult to obtain in the case of greyhounds.