Chloroplast expression systems are promising platforms for recombinant protein production.  Chloroplasts provide an excellent environment for proper folding of heterologous eukaryotic proteins.  Transgenic microalgae that grow non-photosynthetically provide a protected intracellular space that is non-essential to cell growth.  We have recently demonstrated the ability of C. reinhardtii chloroplasts to express a unique class of proteins, consisting of antibody molecules genetically linked to Pseudomonas exotoxin A.  These proteins, termed �immunotoxins�, have the ability to target cancerous cells and deliver the cell-killing toxin.In this presentation, we report our efforts in expressing and purifying two immunotoxins.  Challenges related to downstream processing and recovery of recombinant fusion proteins will be presented.  Specifically, we will discuss optimal conditions for extraction and clarification of algal tissue and the required pretreatment methods to address extract components that are detrimental to immunotoxin activity and purification efficiency.

Expression and recovery of immunotoxins from <i>Chlamydomonas reinhardtii</i> chloroplast

243rd National Meeting (2012)

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Expression and recovery of immunotoxins from Chlamydomonas reinhardtii chloroplast