PHA is an attractive biomaterial for fabrication of novel micro/nano-scaled structures and well-defined surfaces, particularly for medical applications. At our laboratory we fabricate micro/nano-scaled medium-chain-length (mcl) PHA beads. In contrast to standard methods for emulsion-based polymer bead production, detergents are not needed in our process. The mcl-PHA beads can be used for non-specific as well as targeted immobilization of proteins. The binding capacity is comparable to similar-sized polystyrene particles used e.g. for antibody immobilization in latex agglutination tests. Similar applications can be envisaged for mcl-PHA latex given a high capacity for antibody adsorption observed in our studies.
Furthermore, we developed a novel system for targeted immobilization of proteins onto mcl-PHA by using the predicted mcl-PHA-binding domain of an extracellular PHA depolymerase (ePhaZMCL). Mcl-PHA beads sensitized with antibodies may be used for drug delivery to specific target cells in the human body. The advantages of mcl-PHA carriers would be the avoidance of detergents and a high degree of biocompatibility combined with degradation via (slow) surface erosion. Thus, processed mcl-PHA beads are prime candidates for either non-specific or targeted immobilization of proteins in applications such as drug delivery, protein microarrays and protein purification.
In the course of the study we also established a high yield expression system for recombinant, active and secreted ePhaZMCL, facilitating its use for microscale structuring of mcl-PHA surfaces and other biocatalytic applications. |
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