The present invention provides engineered purine nucleoside phosphorylase (PNP) enzymes, polypeptides having PNP activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. Methods for producing PNP enzymes are also provided. The present invention further provides compositions comprising the PNP enzymes and methods of using the engineered PNP enzymes. The present invention finds particular use in the production of pharmaceutical compounds.

The present invention relates to a method for the high yield production of the anticancer nucleoside clofarabine, the method comprising the preparation of 2-chloroadenosine by enzymatic transglycosylation between 2-chloroadenine and nucleosides, benzoylation, isomerization, sulfonate ester formation, fluorination, and deprotection.

The invention provides a method of determining a predisposition to a condition associated with ATP depletion, such as an ischaemic event, in a subject comprising: a. measuring the concentration of one or more purines in a body fluid of the subject, the purines being selected from adenosine, inosine, hypoxanthine, xanthine and ATP, and b. comparing the measured concentration with a threshold concentration of the one or more purines, wherein the threshold concentration is preferably in the range 2 [micro]M to 8 [micro]M and wherein a measured concentration higher than the threshold concentration indicates the presence of ischaemia.

The present invention provides engineered purine nucleoside phosphorylase (PNP) enzymes, polypeptides having PNP activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. Methods for producing PNP enzymes are also provided. The present invention further provides compositions comprising the PNP enzymes and methods of using the engineered PNP enzymes. The present invention finds particular use in the production of pharmaceutical compounds.

The present invention relates to a method for the high yield production of the anticancer nucleoside clofarabine, the method comprising the preparation of 2-chloroadenosine by enzymatic transglycosylation between 2-chloroadenine and nucleosides, benzoylation, isomerization, sulfonate ester formation, fluorination, and deprotection.

The disclosure provides for recombinantly modified Zika-based vectors that can be used in gene therapy applications.

This disclosure relates to lipid nanoparticles comprising nucleic acids encoding therapeutic proteins and uses in treating diseases such as cancer. In certain embodiments, this disclosure relates to methods of treating cancer or initiating, enhancing, or prolonging an anti-tumor response in a subject in need thereof comprising administering to the subject an effective amount of lipid nanoparticles as reported herein comprising a vector or nucleic acid encoding peptide based anticancer agent.