Date of Award


Degree Type


Degree Name

Master of Engineering Science


Chemical and Biochemical Engineering


Dr. Argyrios Margaritis


Recombinant human erythropoietin (rhEPO) is a glycoprotein, which is produced commercially from Chinese hamster ovary (CHO) cells. It is used for chronic therapy of renal anemia and chemotherapy-induced anemia in cancer patients. Recent evidence suggests that rh-EPO exerts tissue protective effects via multiple mechanisms which include inhibition of apoptosis, promotion of angiogenesis and decreased inflammation. After Intra Venus (I.V.) injection, the blood concentration of rh-EPO rapidly decreases due to proteolysis with a short half-life of 8.5 h, which makes treatment expensive. It is desirable to develop an encapsulation method which will provide a controlled release of rh-EPO and maintain the desirable concentration levels in the blood for longer time. Nanoparticles encapsulated rh-EPO can also be used for direct injection to specific tissues/organs, where sustainable concentration of rh-EPO are desirable

In this thesis, we report the production of biopolymer nanoparticles (BNP) with the following methods: (a) emulsification-evaporation (b) ionotropic gelation of the biopolymer chitosan with tripolyphosphate (TPP). The nanoparticle size distribution in aqueous solution was measured using Dynamic Light Scattering system. Three different methods were used and compared for the measurement of rh-EPO concentration in PBS aqueous solution, namely, Enzyme Linked Immunosorbent Assay (ELISA), High Performance Liquid Chromatography (HPLC), and Fluorometry. A series of experiments were performed on the encapsulation of rh-EPO in chitosan biopolymer nanopartciles using the iontropic gelation method with TPP. Rates of release of rh-EPO encapsulated in chitosan-TPP nanoparticles were measured in Phosphate Buffered Saline solution (PBS).

The concentration versus time release data were used with second order diffusion equation to estimate the effective diffusivity of rh-EPO in chitosan-TPP nanoparticles. It was concluded that the chitosan-TPP biopolymer nanoparticles were the best method for the encapsulation of rh-EPO.



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