Methods for the detection, enumeration and analysis of circulating tumor cells expressing insulin-like growth factor-1 receptors (IGF-1R) are disclosed. These methods are useful for cancer screening and staging, development of treatment regimens, and for monitoring for treatment responses, cancer recurrence or the like. Test kits that facilitate the detection, enumeration and analysis of such circulating tumor cells are also provided.

Methods for the detection, enumeration and analysis of circulating tumor cells expressing insulin-like growth factor-1 receptors (IGF-1R) are disclosed. These methods are useful for cancer screening and staging, development of treatment regimens, and for monitoring for treatment responses, cancer recurrence or the like. Test kits that facilitate the detection, enumeration and analysis of such circulating tumor cells are also provided.

A two-step chromatography purification scheme is described which selectively captures and isolates the genome-containing rAAV vector particles from the clarified, concentrated supernatant of a rAAV production cell culture. The process utilizes an affinity capture method performed at a high salt concentration followed by an anion exchange resin method performed at high pH to provide rAAV vector particles which are substantially free of rAAV intermediates.

A two-step chromatography purification scheme is described which selectively captures and isolates the genome-containing rAAV vector particles from the clarified, concentrated supernatant of a rAAV production cell culture. The process utilizes an affinity capture method performed at a high salt concentration followed by an anion exchange resin method performed at high pH to provide rAAV vector particles which are substantially free of rAAV intermediates.

A method for reducing a contaminating DNA content of a preparation containing AAV capsids and contaminating DNA, comprising the steps of a) Performing an extraction of DNA with a solid phase bearing positive charges at its surface said solid phase is contacted with the preparation at a pH of 7.0±1.0, and a salt concentration of 10 mM to 200 mM yielding a first fraction, (b) Diafiltering the first fraction by a first tangential flow filtration to obtain a second fraction, (c) Treating the second fraction with DNase, (d) Diafiltering the DNase treated second fraction obtained by step c) by a second tangential flow, (e) filtration to a buffer with pH of 7.0±1.0, and a salt concentration of 10 mM to 20 mM to yield a third fraction, and optionally (f) Concentrating the third fraction by tangential flow filtration before supplemental chromatography.

In accordance with the invention, provided herein are methods for purifying recombinant adeno-associated (rAAV) vector particles.

Methods for preparing CD7-negative, CD3-positive T cells, which optionally express a chimeric antigen receptor, are provided as is a method of using such cells in a method for treating cancer, in particular a CD7+ cancer. In one aspect, the invention provides a method for preparing a population of CD7-negative, CD3-positive T cells by (a) performing a first selection by depleting, from a population of primary immune cells, cells that express CD7 thereby generating a population of CD7-negative cells; (b) performing a second selection by enriching, from the population of CD7-negative cells, T cells that express CD3 thereby generating a population of CD7-negative and CD3-positive T cells, and (c) incubating the population of CD7-negative and CD3-positive T cells in a culture vessel under stimulating conditions, thereby generating stimulated CD7-negative, CD3-positive T cells.

Site-specific modifications of proteins are desirable in biotechnological applications such as biopharmaceuticals, immunotherapy, vaccines, and are useful in chemical biology. Gluconoylation is a non-enzymatic, covalent, post-translational modification commonly observed on N-terminal His-Tags bearing proteins. We synthesized glucono-1,5-lactone derivatives, including azido variants for selective acylation. High yield acylation is achieved by simply mixing derivatives with target protein amidst diverse conditions of temperatures, aqueous buffers, excipients, or complex cell lysate.

A method for separating cells in a biofluid includes pretreating the biofluid by introducing a predetermined amount of a cocktail of antibodies, flowing the pretreated biofluid through a microfluidic separation channel, and applying acoustic energy to the pretreated biofluid within the microfluidic separation channel. A system for microfluidic cell separation, capable of separating target cells from non-target cells in a biofluid includes at least one microfluidic separation channel, a source of biofluid, a source of an additive including the cocktail of antibodies, and at least one acoustic transducer coupled to the microfluidic separation channel. A kit for microfluidic cell separation is also disclosed. A method of facilitating separation of cells is also disclosed.

This present invention is directed to variable affinity chromatography apparatus and methods for using the same. In particular, the polarity of the stationary phase or the mobile phase is modulated using an external stimulus. Exemplary external stimulus that can be used in the invention include, but are not limited to, electric field, electromagnetic radiation including UV, Vis, and infrared wavelengths, as well other stimuli that are known to one skilled in the art. Generally, any external stimulation that changes the polarity of a stimulus responsive material can be used. One particular embodiment of the invention provides a chromatography apparatus comprising: (i) a chromatography column having a stationary-phase separation medium contained therein; (ii) an external stimulus generator operatively connected to said chromatography column; and (iii) a chromatography mobile-phase, wherein at least one of said stationary-phase separation medium and said chromatography mobile-phase comprises a stimulus responsive material that adopts a different configuration based on the absence or the presence of said external stimulus, wherein different configurations of said stimulus responsive material results in a different stationary or mobile phase affinity, and wherein said external stimulus is selected from the group consisting of electric field, electromagnetic radiation, and a combination thereof.