Protein Production Core

Dana-Farber Cancer Inst, Boston MA

Investigators

Linked publications & trials

Abstract

ABSTRACT – CORE B The T cell receptor (TCR) enables T cell response to infections, pre-malignant or malignant cellular transformation, or physical damage in higher organisms. Due to sophisticated regulation of development and activation, T cells are able to discern minute differences in surface-presented antigen peptide-major histocompatibility complex (pMHC) molecules, resulting in elimination of unwanted insults while avoiding damage to healthy cells or tissues. A major contributor to this selectivity is the mechanosensor function of the αβTCR and its obligatory developmental precursor, the preTCR. The overall goal of this Program Project is to define the molecular mechanisms responsible for the mechanobiology of the αβTCR through convergent single molecule (SM), structural, and biological investigations. Core B, the Protein Production Core, will support this function by producing proteins and coordinating resources to facilitate such investigations seamlessly. First, Core B will support Projects 1 and 2 with protein production appropriate in scale to each technology including SM, single cell, NMR, and X-ray crystallography. The range of proteins to be produced include TCRαβs, preTCRs, biotinylated and native pMHCs, truncated pMHCs, monoclonal antibodies (mAbs), and antibody antigen binding fragments (Fabs). Core B will work in tandem with Projects 1, 2 and 3 and Core C to design mutants, novel constructs and novel production workflows. Second, Core B will continue to serve as a central repository of DNA sequences and constructs to assist in the transfer of knowledge between projects. Standard and novel vectors designed previously within the Program Project will be available as will an array of optimized cloning protocols. Third, Core B will work closely with Core C and Projects 1, 2 and 3 for the use of specific labeling techniques, including those pioneered within the Program Project. By combining these critical functions, Core B will be a central hub for the transfer of information and material for the entire Program Project.