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      Size Exclusion PEGylation Reaction Chromatography Modelling

      Kapadi, Ajith Nayak
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      Kapadi, A. N. (2006). Size Exclusion PEGylation Reaction Chromatography Modelling (Thesis, Master of Engineering (ME)). The University of Waikato, Hamilton, New Zealand. Retrieved from https://hdl.handle.net/10289/2504
      Permanent Research Commons link: https://hdl.handle.net/10289/2504
      Abstract
      Size exclusion PEGylation reaction chromatography was investigated using a model

      developed by Fee (2005). Column dispersion was neglected and the PEGylation

      reaction was modelled as second order. The model allowed up to four PEG groups to

      be attached to a protein and accounted for succinic acid hydrolysis from activated

      PEG. The model was adapted to simulate a-lactalbumin PEGylation and succinic

      acid hydrolysis from activated PEG in a batch stirred tank so rate parameters from

      stirred tank kinetic experiments could be obtained and the model verified. The model

      was solved using finite differences and simulations run in Matlab. The effect of

      reaction parameters such as timing, length and concentration of PEG and protein

      injection, reaction rates, and model resolution on model simulation results was

      explored.

      In the size exclusion PEGylation simulations it was found that increasing protein

      concentration increased MonoPEG concentrations and increased the ratio of

      MonoPEG to starting protein feed concentration. Increasing PEG pulse length and

      starting PEG concentration initially increased MonoPEG concentration and product

      ratio until all protein had been PEGylated at which point MonoPEG concentration the

      product ratio levelled out. Increasing PEG hydrolysis rates did not affect the amount

      of MonoPEG produced but reduced the activated PEG concentration and increased

      succinic acid concentration. Optimal conditions for producing MonoPEG were found

      to be equal concentrations of PEG and protein, with the PEG injection length twice as

      long as the protein injection, and the PEG injection done immediately after the protein

      injection.
      Date
      2006
      Type
      Thesis
      Degree Name
      Master of Engineering (ME)
      Publisher
      The University of Waikato
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