Cell Line Development for Biomanufacturing
This course provides the basic and advanced principles of genetic engineering in yeast and mammalian cells for the overproduction of a protein of interest. Students will use classical and novel strategies to establish a stable Chinese hamster ovary (CHO) cell line based on the industrially relevant strain, DG44 (DHFR system). Students will also generate a yeast expression system based on Pichia pastoris for the production of the same protein.
For the generation of the mammalian cell line, tasks include the optimization of DNA sequence of the gene of interest, design of the expression vectors, transfection of CHO cells, selection for transfected cells, intracellular amplification of the gene of interest, and clone selection using a dilution cloning technique. Through experimentation, students are exposed to several cell culture techniques such as aseptic techniques, microscopy, growth and sub-culture of adherent and suspension CHO cells, determination of metabolites concentration, and cell banking. Students will use other techniques for the transformation, clone selection and gene amplification to generate the yeast Pichia pastoris cell line.
Techniques such as SDS-PAGE, Western blot, and ELISA are used for the detection and quantification of the active recombinant protein.