Antibodies and Antibody Mimics in Binding Assays

Research output: ThesisDoctoral Thesis (compilation)

Abstract

This thesis presents development of new analytical methods using antibodies and molecularly imprinted polymers (MIPs) in binding assays:

1) The high specificity of antibodies and the wide variety of efficient labels make immunoassay one of the most popular analytical techniques today. Enzyme-linked immunosorbent assay (ELISA) is most widely used, since the enzyme label can be used as a catalyst to provide very high signal amplification. However, non-specific binding of proteins on surfaces may cause erroneous results. This thesis exploits the use of materials such as dextrans and dendrimers to modify surfaces, which are commonly used in ELISA, to reduce the interference of non-specific binding. This was realized by appropriate covalent attachment of the modifiers to the solid supports resulting in remarkable improvements in assay sensitivity and selectivity.

2) Due to the high physical and chemical stabilities, MIPs are increasingly used as alternatives to antibodies as selective binding materials. In general, MIPs are prepared by template-directed polymerization of functional monomers and cross-linkers. During the polymerization, the template guides the assembly of functional monomers in the resulting cross-linked polymer. Upon removal of the template, the polymer retains specific binding sites that can selectively rebind the original template, as well as structurally related molecules such as analyte-enzyme conjugates. In this thesis microspheric MIPs prepared by a precipitation polymerisation have been employed. The molecularly imprinted microspheres displayed favourable binding characteristics and were successfully used in aqueous environments. Novel applications of the MIPs in ELISA and scintillation proximity assay (SPA) were demonstrated. Assays for small molecules such as 2,4-dichlorophenoxyacetic acid and (S)-propranolol using microspheric MIPs proved highly selective and sensitive.

Details

Authors
  • Ioana Wärnmark
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biochemistry and Molecular Biology

Keywords

  • Polymerteknik, biopolymers, ELISA, antibody, molecularly imprinted polymer, Biochemistry, Biokemi, Metabolism, metabolism, Biokemisk teknik, Polymer technology, Biochemical technology
Original languageEnglish
QualificationDoctor
Awarding Institution
Supervisors/Assistant supervisor
  • [unknown], [unknown], Supervisor, External person
Award date2002 May 8
Publisher
  • Ioana Wärnmark, Pure and Applied Biochemistry, Chemical Center, Lund University
Print ISBNs91-628-5222-1
Publication statusPublished - 2002
Publication categoryResearch

Bibliographic note

Defence details Date: 2002-05-08 Time: 10:15 Place: Hörsal B at Chemical Center External reviewer(s) Name: Scheller, Frieder Title: Professor Affiliation: Analytical Biochemistry, Potsdam University, Golm, Germany --- Article: Dextran-modified surface for highly sensitive chemiluminescent ELISAIoana Surugiu, Estera S. Dey, Juraj Svitel, Simona Pirvutoiu and Bengt DanielssonAnalyst, 126,1633-1635 (2001) Article: Sensitive chemiluminescent ELISA for SimazineIoana Wärnmark-Surugiu, Igor Budashov, Lei Ye and Bengt DanielssonSubmitted for publication Article: Enhanced detection of 2,4-dichlorophenoxyacetic acid using a dendrimer-coated Biacore chipIoana Wärnmark-Surugiu, Estera S.Dey and Bengt DanielssonSubmitted for publication Article: An enzyme-linked molecularly imprinted sorbent assayIoana Surugiu, Lei Ye, Ecevit Yilmaz, Anatoli Dzgoev, Bengt Danielsson, Klaus Mosbach and Karsten HauptAnalyst, 125,13-16 (2000) Article: Chemiluminescence imaging ELISA using an imprinted polymer as the recognition element instead of an antibodyIoana Surugiu, Bengt Danielsson, Lei Ye, Klaus Mosbach and Karsten HauptAnalytical Chemistry, 73, 487-491 (2001) Article: Development of a flow injection capillary chemiluminescent ELISA using an imprinted polymer instead of the antibodyIoana Surugiu, Juraj Svitel, Lei Ye, Karsten Haupt and Bengt DanielssonAnalytical Chemistry, 73, 4388-4392 (2001) Article: Scintillation proximity assay using molecularly imprinted microspheresLei Ye, Ioana Surugiu and Karsten HauptAnalytical Chemistry, 74, 959-964 (2002)