Materials and methods for use of constrained cohydration agents in the purification of biological materials such as antibodies, viruses, cells, and cellular organelles in connection with convective chromatography, fluidized bed or co-precipitation applications.

Described herein are hydrogel compositions comprising cross-linked bacteriophages. The hydrogels are typically bioactive, degradable, for example biodegradable, self-healing, fluorescent, for example autofluorescent, and/or birefringent. The hydrogels described herein may be used as therapeutics or diagnostics, as scaffolds for material synthesis, as catalysts, as membranes or filters, or as biosensor substrates, for example.

The present invention relates to a plasmid-based CTX phage replication system and Vibrio cholerae strain that can be infected by CTX phage and can be used for cholera toxin production. More particularly, the present invention provides a Vibrio cholera variant strain, which expresses a toxT protein in which tyrosine at position 139 is substituted by phenylalanine through the point mutation of a toxT gene using a plasmid-based CTX phage replication system, and is used as a receptor strain which can improve CTX phage infection efficiency and allows a plurality of CTX prophages to simultaneously infect the strain and to be inserted into the chromosome thereof, which the consequent provision of the effect of increasing the production yield of a cholera toxin.

The present invention relates to methods and systems for the directed evolution of macromolecules. The methods involve increasing the mutation rate of an evolving organism comprising a gene of interest that encodes a gene product lacking a desired activity, whereby a mutated gene of interest is produced that encodes an evolved gene product comprising the desired activity and causing a suppression of mutagenesis. The systems comprise an evolving organism comprising a gene of interest encoding a gene product to be evolved, a host organism, and optionally, a lagoon, a cellstat and/or a suitable growth medium.

The present invention relates to a plasmid-based CTX phage replication system and Vibrio cholerae strain that can be infected by CTX phage and can be used for cholera toxin production. More particularly, the present invention provides a Vibrio cholera variant strain, which expresses a toxT protein in which tyrosine at position 139 is substituted by phenylalanine through the point mutation of a toxT gene using a plasmid-based CTX phage replication system, and is used as a receptor strain which can improve CTX phage infection efficiency and allows a plurality of CTX prophages to simultaneously infect the strain and to be inserted into the chromosome thereof, which the consequent provision of the effect of increasing the production yield of a cholera toxin.

This invention describes the production and properties of a pMHC Multiplexer. The pMHC Multiplexer is a spatially limited composition of two different molecules, an encoding molecule (i.e. an RNA or DNA molecule), and an encoded peptide, where said encoded peptide is encoded by said encoding molecule. Furthermore, the peptide is complexed to a MHC complex and thus is part of a pMHC complex. A preferred embodiment of the invention describes the production and properties of an example pMHC Multiplexer that is a phage particle carrying on its surface a number of identical pMHC complexes, where the peptide of the pMHC complexes is encoded by the DNA contained within the phage particle, and where a covalent or non-covalent bond links a phage coat protein with a pMHC complex and/or a pMHC Multimer. Another preferred embodiment of the invention describes the production and properties of an example pMHC Multiplexer that is a eukaryotic cell carrying on its surface a number of identical pMHC complexes, where the peptide of the pMHC complexes is encoded by the DNA contained within the cell. Yet another preferred embodiment of the invention describes the production and properties of an example pMHC Multiplexer where the encoding molecule is a DNA or RNA, and where the binding of pMHC Multiplexer to T cell receptor (TCR) can be detected by PCR-based analysis. Yet another preferred embodiment of the invention describes the production and properties of an example pMHC Multiplexer that comprises one or more identical pMHC complexes, where the encoding molecule is directly linked to at least one peptide (p) of one of the pMHC complexes, and thus, the peptide (p) of the pMHC complex(es) is encoded by said encoding molecule directly linked to it.

The present invention relates to methods and systems for the directed evolution of macromolecules. The methods involve increasing the mutation rate of an evolving organism comprising a gene of interest that encodes a gene product lacking a desired activity, whereby a mutated gene of interest is produced that encodes an evolved gene product comprising the desired activity and causing a suppression of mutagenesis. The systems comprise an evolving organism comprising a gene of interest encoding a gene product to be evolved, a host organism, and optionally, a lagoon, a cellstat and/or a suitable growth medium.

A method of purifying a sample that includes a desired virus includes the steps of (i) providing a packed chromatographic column having negatively charged porous particles, (ii) equilibrating the column to the conditions to which the desired virus in the sample is to elute, (iii) contacting the sample with the packed chromatographic column such that the sample volume applied to the packed chromatographic column is less than or equal to the interparticle space of the negatively charged porous particles within the packed chromatographic column, (iv) eluting the desired virus from the packed chromatographic column, where the desired virus is in a purer state and in the conditions to which the packed chromatographic column was equilibrated.

Materials and methods for use of constrained cohydration agents in the purification of biological materials such as antibodies, viruses, cells, and cellular organelles in connection with convective chromatography, fluidized bed or co-precipitation applications.

Described herein is a high-throughput method of synthesizing biofunctional microparticles. In aspects, the method comprises casting biofunctional microparticle precursors onto a microporous template to form microparticles, wherein the template comprises a removable film; and removing the film to liberate the microparticles. Also described herein is a sprayable microgel and related methods.