Disclosed herein are polypeptides having nuclease activity. Some of the polypeptides having nuclease activity were generated by an improved gene site mutagenesis (“GSSM”) method or the tailored multi-site combinatorial assembly (“TMCA”) method. Also disclosed are compositions and kits comprising the polypeptides having nuclease activity, and methods for making and using these polypeptides, compositions and kits.

Disclosed is an engineered thermolabile mutant of Serratia Marcescens Nuclease. In the same buffer as used for optimal enzyme activity, after 60 C. for 20 min, while the wild type retained 12.5% activity, the thermolabile mutant R146D/D156R/D229R/D245R (SEQ ID NO: 12, without the first 21 amino acids, which are a signal peptide) retained only 0.39% activity. Heat inactivation of the mutant, when it is used for protein purification, can be used after a period when substantial DNA in a protein has been degraded, but before unwanted degradation takes place.

The present invention provides methods for detecting a target nucleic acid in a sample by, for example, incubating the target nucleic acid with a detection probe containing a nucleic acid sequence complementary to at least a portion of the target nucleic acid and a nuclease enzyme that specifically cleaves double-stranded nucleic acids. Hybridization between the detection probe and the target nucleic acid thereby leads to cleavage of the detection probe, releasing a portion of the probe attached to a detectable agent. The portions of the digested probes attached to the detectable agent can be separated from unbound probe and detected in order to determine the presence of the target nucleic acid in the sample. Thus, the invention enables rapid and accurate analysis of a sample for the presence of desired nucleic acid biomarkers.

A fabric softener composition comprising a quaternary ammonium ester fabric softener compound and an enzyme selected from specific nuclease enzymes, galactanase enzymes and mannanase enzymes. Also, methods of treating a fabric comprising a laundering step, optional rinsing steps and a rinse-treatment step in which the fabric is treated with an aqueous rinse liquor comprising the composition.

The invention relates to a method for purifying an adenovirus comprising (a) providing a liquid sample comprising adenovirus, (b) clarifying said sample by depth filtration, (c) performing anion exchange chromatography comprising the steps of (i) directly applying the clarified sample of (b) to an anion exchange column, (ii) eluting adenovirus from the anion exchange column to provide an eluate. The invention also relates to adenovirus produced from said methods, and to compositions comprising same.

A method for purifying exosomes from a cell culture medium comprising combining at least three (03) treating stages of a sample containing exosomes in the following specific order: (A) filtering the sample containing exosomes by tangential flow filtration (TFF) to obtain a TFF-filtered sample; (B) centrifuging the TFF-filtered sample by sucrose cushion centrifugation (SCC) to obtain a SCC-centrifuged sample; and (C) purifying the SCC-centrifuged sample by bind-elute size exclusion chromatography (BE-SEC) to obtain a purified exosomes. The present invention provides a comprehensive and efficient approach for purifying exosomes from a cell culture medium, facilitating using exosomes in various biomedical applications.

Either wild type or mutant Serratia marcescens Nuclease (SMN) is engineered to display a C-terminal Chitin Binding Domain (CBD-tag), followed, at the C-terminus side of the CBD-tag, by a poly-histidine tag (His-tag), where the His-tag is preferably a 6-mer and preferably is preceded by a Gly-Ser linker, thereby generating a recombinant SMN protein that retains dual affinity tags (a CBD-tag and a His-tag) at the C-terminus, to make it easily removed from a reaction solution following digestion of nucleic acids in the reaction mixture. It can also be used for binding SMN to a solid support for use in nucleic acid digestion in a sample contacted with the solid support.

A fabric softener composition comprising a quaternary ammonium ester fabric softener compound and an enzyme selected from specific nuclease enzymes, galactanase enzymes and mannanase enzymes. Also, methods of treating a fabric comprising a laundering step, optional rinsing steps and a rinse-treatment step in which the fabric is treated with an aqueous rinse liquor comprising the composition.

The objective of the present invention is to provide a method capable of efficiently removing an impurity from an aqueous solution or a suspension comprising an antibody or an antibody-like molecule and the impurity. The method for purifying an antibody or an antibody-like molecule according to the present invention is characterized in treating an aqueous solution or a suspension comprising the antibody or the antibody-like molecule and an impurity with a water-insoluble inorganic compound, wherein the water-insoluble inorganic compound comprises one or more elements selected from magnesium, calcium and aluminum.

A system for an ocular gene therapy is provided. The system includes one or more optimized transgenes. The one or more optimized transgenes include pAAV.CMV.CodOpt.RPE65 and pAAV.CMV.Kozak.RPE65. The optimized transgenes pAAV.CMV.CodOpt.RPE65 and pAAV.CMV.Kozak.RPE65 have shown to exhibit enhanced RPE65 gene expression when compared to wild type RPE65 gene transfer in suitable models. These optimized genes may enhance therapeutic response during LCA2 gene therapy. The present invention also provides a process for preparing the optimized transgene for an ocular gene therapy.