The Recombinant Protein Production Core (rPPC) provides quality controlled recombinant proteins for researchers within the Northwestern Community (WCAS, McCormick, Feinberg) and also serves academic and industry researchers outside of Northwestern University. rPPC operates based on the two service models: (1) a Training model where Northwestern researchers use specialized bioreactor systems and participate in hands-on-training activities and (2) a Production model where staff carry out expression (mg to gm scale) and purification of recombinant or synthetic biologics, including potential therapeutic proteins and peptides, among others. Currently, the main focus of rPPC is to be a user-facility; the facility has parallel bioreactor systems for multiplexed lab-scale cultivation of microbial, insect, and mammalian cells. rPPC also serves as a production facility, providing low-cost recombinant biologics for researchers at Northwestern University.
rPPC’s goal is to transform research at Northwestern University by addressing a variety of unmet needs that can often stall cutting-edge projects. As a centralized protein production center rPPC facilitates the expression of proteins for a broad spectrum of research activities ranging from structure/function studies to producing protein therapeutic candidates such as monoclonal antibodies for in vitro and in vivo pre-clinical studies. Scalable, high-throughput cultivation enables screening mutants for phenotype studies, proteomics, stem cell characterization, media screening, strain and process optimization for the production of biochemicals, e.g. biofuels. High-throughput cell-free protein production interfaces well with HTA to “augment and expand Northwestern University’s community of multidisciplinary researchers focusing on areas of biomedical research relevant to NIH.” Protein purification completes the facility enabling a one-stop shop for researchers to clone, overexpress, and purify relevant biologics.
All prices are at our External Academic rate. For corporate rates, please contact us.
The DNA provided by customer will be expressed in competent E.coli cells. The scaled up E.Coli culture will be grown in shaker flask. DNA will be extracted and purified using plasmid mini - or maxi-prep kit depending on customer's request.
Recombinant protein expression involves transforming bacterial cells with a DNA vector that contains the template and then culturing the cells so that they transcribe and translate the desired protein. The cells are then lysed to extract the expressed protein for subsequent purification. Protein can be expressed in shaker flasks or on a 1L or 5L scale in bioreactors.
Insect cells are cotransfected with wild type baculovirus DNA and transfer vector plasmid DNA containing the foreign gene to be expressed to generate recombinant baculovirus carrying the DNA of interest. Price is per Liter of culture. We use a Bac-to-Bac system, from Invitrogen and Sf9 cells.
Recombinant protein expression involving transfecting cells with a DNA vector that contains the template and then culturing the cells so that they transcribe and translate the desired protein. The cells are then either lysed to extract the expressed protein for downstream processing, or growth medium is collected for secreted protein. Culture growth optimization is possible by using Q Plus Fermenter (1L, up to 3 in parallel).
This service includes the steps immediately following protein expression and immediately preceding protein purification. Downstream processing is used to recover active recombinant protein. It includes cell separation by centrifugation, cell membrane disruption and cell lysis by high pressure homogenizer.
A spectrum of downstream processes are used to recover active recombinant protein: cell separation by centrufugation, cell lysis membrane disruption and cell lysis by high pressure homogenizer, protein separation and fusion tag cleavage. Price is per cell mass from 1L of culture.
The service includes analytical immune detection of a protein in the sample or cell lysates provided by a customer. Using Capillary Electrophoresis, the fully automated ProteinSimple Wes Simple Western size assay system will greatly accelerate protein detection with high precision and quality. This technology is based on nanovolume size-based protein separation that can be used to quantify proteomic profiles of any experimental protein samples, or clinical specimens for both biomarker discovery and diagnostics. Instead of two days you spend just three hours to get accurately quantitated, and reproducible result of 24 samples.
Generation of a hybridoma cell line (secreting antibody) starts with immunization of mice with an antigen of interest (immunogen), and proceeds with fusion of immune splenocytes and myeloma cells to make hybrid cells (hybridomas) that are to be screened to select the ones that produce a specific antibody. Standard screening assays used against the injected antigen (immunogen) include indirect ELISA and Western Blot. Currently rPPC offers only three steps of the hybridoma generation process – immunization, fusion and screening. After two rounds of screening, 96-well plates with growing young hybridoma cells will be transferred to the customer’s laboratory for hybridoma cell preservation by freezing down, and single cell cloning of the selected hybridoma cell line.
Hybridoma culture medium (1L), containing secreted antibody is purified through an automatic liquid chromatography system. The technique separates antibodies on the basis of a chromatography matrix. If a highly concentrated hybridoma supernatant (from our bioreactor) is to be purified, the above price applies to a 50 mL unit.
A plasmid carrying the target DNA provided by customer will be inserted into an expression vector. Cells (bacterial, insect and mammalian) are transformed/transfected with an expression vector carrying the target DNA, encoding the protein of interest.
Recombinant Protein Production Core (Northwestern University) has not received any reviews.
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