An investigation into the large scale separation of equine blood plasma.
Thesis DisciplineChemical Engineering
Degree GrantorUniversity of Canterbury
Degree NameMaster of Engineering
The large-scale separation of proteins from equine blood plasma was investigated, as an attempt to create a process that would separate multiple proteins. This contrasts to the current state, where methods target a single protein at a time. Ion exchange chromatography and ethanol precipitation were both used as techniques, due to their use in human blood plasma fractionation and their versatility as downstream separation methods. The bases of ethanol precipitation and the underlying percentages used were investigated, along with techniques specifically designed to remove albumin. Ion exchange fractions were based on those used in the separation of bovine plasma, with a specific focus on a 20-fold scale-up in column volume. The results indicate that ion exchange chromatography is a feasible separation method. Separation was able to be carried out, with immunoglobulins in particular easily obtainable from this techniques. The overall profile broadly matched that of bovine blood plasma. However, there was considerable amounts of buffer required, along with significantly increased column pressure upon scale-up. These drawbacks can be overcome, but this may restrict the applicability of the process as an initial step due to the expense of buffers and columns that can withstand high pressures. Ethanol precipitations, on the other hand, proved effective in separation and would be easier to scale up for an industrial process, although requiring flame-proof equipment and facilities at large scale. While issues were found with the techniques used to identify the proteins, it was clear that in particular, albumin separation methods from the bovine plasma industry would easily be adaptable to equine blood. From this, a complete process was obtained for use in both existing and new plants. With only three sub-categories of proteins commercialised from equine plasma, the scope for minimising waste protein was severely restricted. In addition, many of the methods of separation are quite expensive, further restricting the number of proteins that could be obtained. However, there is the ability to reduce waste through multiple protein production without major capital investment.