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Preparation and characterization of pretargeting molecules: The antibody BR96-Streptavidin conjugate
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
2010 (English)Independent thesis Advanced level (degree of Master (One Year)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Introduction: In pretargeted radio immunotherapy, one molecule is administrated to bind to the target cell, e.g. a tumour cell, and after clearance another molecule, a radio labeled effecter molecule, which binds to the pretargeting molecule, is administrated.

Aim: In this thesis work methods are tested and validated for conjugation of a monoclonal antibody with streptavidin by two separate conjugation methods. One of the methods involves sulfo-SMCC (sulfo-Succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate) and SATA (S-acetylthioacetic acid) and the other one is based on the Staudinger reaction with NHS-azide and NHS-phosphine. Moreover, preparation of F(ab)2- and Fab-fragments of the monoclonal antibody with immobilized pepsin and papain, respectively,  is evaluated. The plan is that the antibody-streptavidin conjugate shall be used as a pretargeting molecule while radio labelled biotin will be used as an effecter molecule.

Method: The formed conjugates are evaluated by using Agilent Bioanalyzer 2100, by  immunoreactivity investigations, and to some extent by gel filtration using an ÄKTA system.

Result: The sulfo-SMCC reaction results in conjugates with higher immunoreactivity than the  Staudinger based reaction, which seems to be more efficient. The conjugates prepared by the later method might be less functional due to the fast reaction resulting in undetectable large multi-complexes.

Conclusion: Streptavidin-IgG complexes were efficiently formed by both conjugation procedures used. However, it was difficult to characterize the obtained complexes by the utilised methods.

Place, publisher, year, edition, pages
2010.
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:lnu:diva-6461OAI: oai:DiVA.org:lnu-6461DiVA, id: diva2:326462
Uppsok
Physics, Chemistry, Mathematics
Supervisors
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Available from: 2010-09-02 Created: 2010-06-23 Last updated: 2010-09-02Bibliographically approved

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CiteExportLink to record
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Cite
Citation style
  • apa
  • ieee
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