Institutional Research Information Service
UCL Logo
Please report any queries concerning the funding data grouped in the sections named "Externally Awarded" or "Internally Disbursed" (shown on the profile page) to your Research Finance Administrator. Your can find your Research Finance Administrator at https://www.ucl.ac.uk/finance/research/rs-contacts.php by entering your department
Please report any queries concerning the student data shown on the profile page to:

Email: portico-services@ucl.ac.uk

Help Desk: http://www.ucl.ac.uk/ras/portico/helpdesk
Publication Detail
Does PEGylated DI have the potential to be an effective therapeutic for Antiphospholipid Syndrome?
  • Publication Type:
  • Authors:
    McDonnell TCR
  • Pagination:
    1, 330
  • Supervisors:
    Rahman ,Ioannou
  • Awarding institution:
  • Date Submitted:
  • Keywords:
    Antiphospholipid Syndrome, PEGylation, Biological Therapeutic, Lupus Anticoagulant, ELISA, Protein Production, Refolding
  • Addresses:
    Thomas Christopher Richard McDonnell
    University College London
    Division of Medicine
    Rayne Institute, 5 Univeristy Street
    United Kingdom

    Thomas Christopher Richard McDonnell
    3 Springcrofts, Bushey, Watford, Herts
    WD23 3AR
    United Kingdom
Antiphospholipid Syndrome (APS) has 0.3-1% population prevalence and characterised by clinical phenotypes (thrombotic and obstetric) and the presence of anti-phospholipid antibodies (aPL). APS is the leading cause of strokes in patients <50 years old and causes recurrent miscarriage. Current treatment is inadequate and consists of life long non-specific anticoagulant therapy. It has been previously proven that Domain I (DI) of Beta-2-glycoprotein I (β2GPI) harbours the dominant epitope for pathogenic antibodies in APS and administration of a recombinant version of DI can abrogate clotting In an APS mouse model. Despite this, DI requires chemical modification to be suitable for drug administration. This thesis details the production methodology, characterisation and results of chemical conjugation of DI to poly ethylene glycol (PEG). Production remains a difficult hurdle in the production of biological based therapeutics. I increased production to 75 mg/L (from 4-6 mg/L). I then semi-automated the folding process, increasing yields and generated purification protocols yielding a purity of >95% throughout. PEGylation was undertaken to extend half life and reduce immunogenicity. I achieved conjugation yields of ~ 50% mono-PEGylated species (molar yield) with minimal waste Di-PEGylated product. Expression tag was removed and endotoxin removed successfully and prepared for final testing. Activity testing included a novel thrombotic functional test, two competition ELISAs (IgG (n=6) & IgA (n=4) ) and a mouse model. All assays showed a high level of retained activity for the protein. Using a novel functional, thrombotic test (n=4), results suggested a gain of function for WT PEGylated species but not PEGylated mutant DI species. In the IgG ELISAs on average >80% activity was retained with similar seen in the IgA assays. Together, these results suggest that PEGylated DI harbours the potential to be the first specific, targeted therapeutic in APS.
Publication data is maintained in RPS. Visit https://rps.ucl.ac.uk
 More search options
UCL Researchers
Div of Medicine
Dept of Biochemical Engineering
University College London - Gower Street - London - WC1E 6BT Tel:+44 (0)20 7679 2000

© UCL 1999–2011

Search by