Artificial elevation of HDL levels (high density lipoprotein) was identified for its potential in the treatment of acute cardiovascular events in the early 1980s. However the major hurdle in developing HDL compounds is the large amount of drug needed for late stage development and market coverage. Whereas other therapeutic proteins are potent at low doses, effective drug amounts per patient for apolipoprotein-based therapeutics (reconstituted HDL) tend to be in the gram range. Thus a yearly production of greater than 1 ton would be needed.To complicate matters further, apolipoproteins, the major protein constituents of HDL particles, have a very low solubility under native conditions and a strong tendency to bind endotoxins. One obvious solution to resolve these difficulties during purification is the use of denaturants such as guanidinium chloride (GuaCl) and  urea. However working under denaturing conditions limits process scalability and makes mass production impossible due to the unmanageable amounts of chaotropic salts required. To date there are no publications reporting the purification of apolipoproteins under native-like conditions.The goal of this study was to develop an innovative manufacturing process for reconstituted HDL particles which allows scalability to a final yield of many kg per batch. Such a process would reduce the use of denaturants and limit the number of process steps in order to be suitable as a scalable and cost-effective market process.

Downstream Process development of an Apo-A1 fusion protein, a complex API expressed in Escherichia coli

247th National Meeting (2014)

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