Abstract: T cell receptors (TCRs) that have higher affinity for the Survivin antigen are provided. The high affinity TCRs were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the engineered TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Abstract: The use of model T cell receptors (TCRs) as scaffolds for in vitro engineering of novel specificities is provided. TCRs with de novo binding to a specific peptide-major histocompatibility complex (MHC) product can be isolated by: 1) mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants (a library), 2) selection of the library of TCR mutants with the specific peptide-MHC, using a process of directed evolution and a “display” methodology (e.g., yeast, phage, mammalian cell) and the peptide-MHC ligand. The process can be repeated to identify TCR variants with improved affinity for the selecting peptide-MHC ligand.

Abstract: A soluble human single-chain T cell receptor (TCR) having the structure: V?2-L-V? or V?-L-V?2, wherein L is a linker peptide that links V? with V?, V? is a TCR variable ? region, and V?2 is a TCR variable ? region of the family 2 is provided. The provided scTCR is useful for many purposes, including the treatment of cancer, viral diseases and autoimmune diseases.

Abstract: T cell receptors (TCRs) that have specificity for the WT1 antigen are provided. The TCRs include higher affinity TCRs that were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Abstract: T cell receptors (TCRs) that have higher affinity for the Survivin antigen are provided. The high affinity TCRs were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the engineered TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Abstract: The use of model T cell receptors (TCRs) as scaffolds for in vitro engineering of novel specificities is provided. TCRs with de novo binding to a specific peptide-major histocompatibility complex (MHC) product can be isolated by: 1) mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants (a library), 2) selection of the library of TCR mutants with the specific peptide-MHC, using a process of directed evolution and a “display” methodology (e.g., yeast, phage, mammalian cell) and the peptide-MHC ligand. The process can be repeated to identify TCR variants with improved affinity for the selecting peptide-MHC ligand.

Abstract: T cell receptors (TCRs) that have higher affinity for the Survivin antigen are provided. The high affinity TCRs were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the engineered TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Abstract: T cell receptors (TCRs) that have specificity for the WT1 antigen are provided. The TCRs include higher affinity TCRs that were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Abstract: T cell receptors (TCRs) that have higher affinity for the Survivin antigen are provided. The high affinity TCRs were engineered through the generation of mutational libraries of TCRs in a single-chain format, followed by selection for improved stability and affinity on the surface of yeast (i.e. directed evolution). In embodiments, the engineered TCRs can be used in soluble form for targeted delivery in vivo, or as genes introduced into T cells in an adoptive T cell setting.

Abstract: The present invention provides a genetic method of tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering T cell receptor binding affinity and specificity by library screening.

Abstract: T cell receptors (TCRs) that have higher affinity for a ligand than wild type TCRs are provided. These high affinity TCRs are formed by mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants of the T cell receptor protein coding sequence; transforming the T cell receptor mutant coding sequence into yeast cells; inducing expression of the T cell receptor mutant coding sequence on the surface of yeast cells; and selecting those cells expressing T cell receptor mutants that have higher affinity for the peptide/MHC ligand than the wild type T cell receptor protein. The high affinity TCRs can be used in place of an antibody or single chain antibody.

Abstract: The use of model T cell receptors (TCRs) as scaffolds for in vitro engineering of novel specificities is provided. TCRs with de novo binding to a specific peptide-major histocompatibility complex (MHC) product can be isolated by: 1) mutagenizing a T cell receptor protein coding sequence to generate a variegated population of mutants (a library), 2) selection of the library of TCR mutants with the specific peptide-MHC, using a process of directed evolution and a “display” methodology (e.g., yeast, phage, mammalian cell) and the peptide-MHC ligand. The process can be repeated to identify TCR variants with improved affinity for the selecting peptide-MHC ligand.

Abstract: An illuminator is described which may be used with large inspection areas and which provides a dark field illumination pattern that is spatially uniform, illuminates from consistent angles, has high efficiency, and is smaller than existing solutions. A light pipe has a first end proximate an object to be illuminated and a second end opposite the first end and spaced from the first end. The light pipe also has at least one reflective sidewall. The first end of the light pipe includes an exit aperture and the second end has at least one opening to allow at least one image acquisition device to view the surface therethrough. At least one light source is configured to provide illumination in the light pipe. The object is illuminated by the first end of the light pipe by illumination at a selected elevation angle and substantially all azimuth angles.

Abstract: An optical inspection system is provided for inspecting a workpiece including a feature to be inspected. The system includes a workpiece transport configured to transport the workpiece in a nonstop manner. An illuminator is configured to provide a first strobed illumination field type and a second strobed illumination field type. The illuminator includes a light pipe having a first end proximate the feature, and a second end opposite the first end and spaced from the first end. The light pipe also has at least one reflective sidewall. The first end has an exit aperture and the second end has at least one second end aperture to provide a view of the feature therethrough. An array of cameras is configured to digitally image the feature. The array of cameras is configured to generate a first plurality of images of the feature with the first illumination field and a second plurality of images of the feature with the second illumination field.

Abstract: The present invention provides a genetic method of tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering T cell receptor binding affinity and specificity by library screening.

Abstract: The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. The outer wall of each yeast cell can display approximately 104 protein agglutinins. The native agglutinins serve as specific adhesion contacts to fuse yeast cells of opposite mating type during mating. In effect, yeast has evolved a platform for protein-protein binding without steric hindrance from cell wall components.

Abstract: A soluble human single-chain T cell receptor (TCR) having the structure: V?2-L-V? or V?-L-V?2, wherein L is a linker peptide that links V? with V?, V? is a TCR variable ? region, and V?2 is a TCR variable ? region of the family 2 is provided. The provided scTCR is useful for many purposes, including the treatment of cancer, viral diseases and autoimmune diseases.

Abstract: Provided are high affinity T cell receptor variable regions that are useful for treating diseases caused by superantigens including atopic dermatitis, pneumonia and delayed wound healing. The variable regions contain mutants that result in high affinity binding to the superantigen.

Abstract: Stabilized variable regions of the T cell receptor and methods of making the same using directed evolution through yeast display are provided. In one embodiment, the variable region is variable beta. In one embodiment, the stabilized T cell receptor variable regions have high affinity for a superantigen, such as TSST-1 or SEB. These T cell receptor variable regions are useful as therapeutics.

Abstract: An optical inspection system is provided for inspecting a workpiece including a feature to be inspected. The system includes a workpiece transport configured to transport the workpiece in a nonstop manner. An illuminator is configured to provide a first strobed illumination field type and a second strobed illumination field type. The illuminator includes a light pipe having a first end proximate the feature, and a second end opposite the first end and spaced from the first end. The light pipe also has at least one reflective sidewall. The first end has an exit aperture and the second end has at least one second end aperture to provide a view of the feature therethrough. An array of cameras is configured to digitally image the feature. The array of cameras is configured to generate a first plurality of images of the feature with the first illumination field and a second plurality of images of the feature with the second illumination field.