The present disclosure provides regulatable biocircuit systems, effector modules and compositions for cancer immunotherapy. Methods for inducing anti-cancer immune responses in a subject are also provided.

The present disclosure relates to mutant polypeptides derived from Escherichia coli dihydrofolate reductase (DHFR) which can be fused to a polypeptide of interest for efficient conditional modulation of its activity. Also disclosed are polynucleotides encoding such mutant polypeptides, vectors comprising such polynucleotides, and the use of such polypeptides, polynucleotides and vectors for treating a disorder.

The disclosure provides methods for carrying out Real Time Cellular Thermal Shift Assays (RT-CETSA). Also provided are molecular constructs and protein constructs for use in such assays and devices suitable for carrying out such assays.

The present invention includes an engineered cell comprising a chimeric antigen receptor (CAR) further comprising a nucleic acid molecule comprising a suicide gene comprising a ligand binding domain and a suicide domain wherein the ligand binding domain is capable of binding to radiolabeled tracer or a small molecule suicide switch. This invention also includes methods for inducing apoptosis of an engineered cell, methods for assessing the efficacy or toxicity of an adoptive cell therapy in a subject, methods for detecting the quantity of engineered T cells in a subject, methods for monitoring an immunotherapy treatment in a subject and methods of imaging engineered T cells in a subject. In some embodiments, the imaging is performed via Positron Emission Topography (PET). This invention further includes a chemical inducer of dimerization (CID), wherein the CID is a Bis-Trimethoprim (Bis-TMP).

The present invention relates to an optimal medium for growing a cell line auxotrophic for tetrahydrofolate (THF) and producing a desired material in the cell with high efficiency. In particular, the present invention provides a method for enhancing cell growth by adding tetrahydrofolate (THF), or a precursor or derivative thereof into a chemical composition cell medium.

This disclosure relates to a virus-like particle in which a small molecule-protein complex is entrapped, ensuring the formation of the small molecule-protein complex under physiological conditions, while protecting the small molecule-protein complex during purification and identification. The disclosure further relates to the use of such virus-like particle for the isolation and identification of small molecule-protein complexes.

The invention pertains to an expression vector or a combination of at least two expression vectors comprising at least (a) a polynucleotide encoding a product of interest or an insertion site for incorporating a polynucleotide encoding a product of interest; (b) a polynucleotide encoding a first selectable marker (sm I); (c) a polynucleotide encoding a second selectable marker (sm II), which is different from the first selectable marker (sm I),
wherein the activity of the selectable marker (sm I) or (sm II) is at least partially influenced by the activity of the other selectable marker and wherein the selectable markers (sm I) and (sm II) are involved in the folate metabolism. Also provided are suitable host cells, selection methods and methods for producing polypeptides with high yield.

An expression system for expressing a protein comprising: a eukaryotic host cell carrying a dihydrofolate reductase (DHFR) deficiency; and an expression vector, the expression vector encoding the human growth hormone gene; a expression vector, the expression vector comprising: a eukaryotic selectable marker including a minimal SV 40 early promoter driving expression of a sequence encoding dihydrofolate reductase for complementing the DHFR deficiency in the host cell; a prokaryotic selectable marker conveying Ampicillin resistance to a prokaryotic host cell; a prokaryotic Origin of Replication; a plurality of multiple cloning sites (MCS); and at least one protein expression module comprising: a Simian Vacuolating Virus 40 (SV40) early promoter, inclusive of its 72 bp enhancer repeats; and a rabbit β-globin intron sequence being separable from a SV40 p A sequence by a first multiple cloning site, for receiving a coding sequence and expressing a desired protein therefrom.

The present invention is directed to the cells, compositions and methods for the production of recombinant protein, wherein an f-met group on the 5′-terminus is enzymatically removed. In particular, the invention is directed to a production process for obtaining high levels of soluble recombinant CRM.sub.197 protein from E. coli. Cells preferably contain one or more mutations of disulfide reductase genes, so that disulfide reductase activity is reduced. The invention also relates to purification method for CRM.sub.197 as well as characterization of properly folded CRM.sub.197 protein.

The present invention provides mammalian cell expression vectors that impart to mammalian host cells an ability to produce high levels of foreign gene-derived proteins. A ubiquitously acting chromatin opening element (UCOE) is introduced into an expression vector that has a plasmid DNA integrated into a transcriptional hot spot on the chromosome of a dihydrofolate reductase gene-deficient host cell so that it allows for selection of strains that grow in hypoxanthine-thymidine (hereinafter denoted as HT)-free medium, whereby transformants will produce a protein of interest in increased amounts.