Synthetic fragment antigen-binding (Fab) antibodies are disclosed that bind to an N-terminal activation site of BCL-2-associated X-protein (BAX) and inhibit BAX activation. Also disclosed are methods of using the Fabs for measuring inactive monomeric BAX levels, screening for small molecules that bind to an N-terminal activation site of BAX, inhibiting apoptotic cell death, and predicting the ability of a cancer therapy to promote apoptotic cell death.

Provided are suspension-based cell culture systems and media for the timely and efficient proliferation of human tumor cell clusters from a patient, and related methods of evaluating the potential responsiveness of the tumor cells and the patient to one or more therapeutic agents.

The present invention relates to the field of anastasis, i.e., the process of reversal of apoptosis. More specifically, the present invention provides methods and compositions useful for studying anastasis. In one embodiment, a tracking construct of the present invention comprises Lyn11-NES-ERT2-DEVD-rtTA-3?FLAG-DEVD-ERT2-NES. In another embodiment, a construct comprises Lyn11-NES-DEVD-rtTA-3?FLAG. In a further embodiment, a construct comprises ERT2-DEVD-rtTA-3?FLAG-DEVD-ERT2.

Synthetic fragment antigen-binding (Fab) antibodies are disclosed that bind to an N-terminal activation site of BCL-2-associated X-protein (BAX) and inhibit BAX activation. Also disclosed are methods of using the Fabs for measuring inactive monomeric BAX levels, screening for small molecules that bind to an N-terminal activation site of BAX, inhibiting apoptotic cell death, and predicting the ability of a cancer therapy to promote apoptotic cell death.

Molecular probe suitable for quantification of caspase activity in vivo using positron emission tomography (PET). Embodiments of the present invention can detect apoptosis in tumors and as a novel, potentially translatable biomarker for predicting response to personalized medicine.

Provided herein are methods for testing proliferative responses of a drug on patient-derived tumor cells; the method comprising obtaining cells from biopsy or tumor resection material; culturing the cells on a 3D extracellular matrix (ECM); treating the cells in ECM with a drug; subjecting the treated cells to high-content (HC) imaging; and evaluating the HC imaging of the treated cells; thereby testing the proliferative responses of the drug on the patient-derived tumor cells. In some embodiments, the methods disclosed herein comprise obtaining cells from biopsy or tumor resection material; xenografting the cells into an animal model (patient-derived xenograft; PDX) for tumor formation; and obtaining tumor cells from the animal.

Cytotoxic factors having use in modulating cell death, and their use in methods of treating necrosis or apoptosis-related conditions are disclosed. The invention also relates to methods for identifying active agents useful in treating conditions related to cell death. The present inventors have found that different pathogens produce different cytotoxic factor(s) having anticancer activity. The substantially pure cytotoxic factors can be used in a method of treating an infectious disease or a cancer.

Provided are suspension-based cell culture systems and media for the timely and efficient proliferation of human tumor cell clusters from a patient, and related methods of evaluating the potential responsiveness of the tumor cells and the patient to one or more therapeutic agents.

Provided is a method for determining an effect of a potential active agent on a sample comprising a co-culture comprising a target cellular object, TCO, and one or more further cell types, FCT, wherein the TCO has been labelled with a flourescent live-cell marker having a first emission/excitation profile, and with a luminescent cell-death marker, and optionally the FCT has been labelled with a fluorescent live-cell marker having a second emission/excitation profile different from the first emission/excitation profile, comprising capturing, during an acquisition event using a microscope, a first fluorescent image from the fluorescent live-cell marker having the first emission/excitation profile, and optionally a second fluorescent image from the fluorescent live-cell marker having the second emission/excitation profile, and a luminescence measurement of the sample, and comparing fluorescent images and luminescent measurements between sample contacted with the potential active agent and sample not contacted with the potential active agent, determining from a difference between test datasets and control datasets an effect of the potential active agent.

Provided herein is a method of detecting microtubule-associated protein 1 light chain 3 (LC3B) expression in a subject as a marker for autophagy, the method comprising: obtaining a liquid biological sample from the subject; isolating exosomes from the liquid biological sample; and detecting LC3B expression in the isolated exosomes. Also provided is a method of detecting LC3B expression as a marker for autophagy in a subject suffering from a solid malignancy, comprising: obtaining a peripheral blood sample from the subject; isolating exosomes from the peripheral blood sample; and detecting LC3B expression in the isolated exosomes, wherein the method does not include obtaining a biopsy of the solid malignancy.