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Insights into weak affinity antibody-antigen interactionsStudies using affinity chromatography and optical biosensor

Strandh, Magnus LU (2000)
Abstract (Swedish)
Popular Abstract in Swedish

I cellen styrs allt av biomolekylernas förmåga till igenkänning och växelverkan med varandra. Molekylernas speciella egenskaper bygger upp kroppens alla komplicerade funktioner, till exempel immunförsvar, celldelning och muskelarbete. Interaktionerna mellan biomolekylerna kan vara av olika karaktär; de beskrivs som starka eller svaga (affinitet), långsamma eller snabba (kinetik).



Det har visat sig att många biologiska mekanismer styrs av svaga interaktioner som, flera tillsammans, skapar igenkänning och stringens. Tack vare att de enskilda interaktionerna är svaga skapas samtidigt en dynamik och förmåga för systemen att snabbt anpassa sig till omgivande förhållanden. Detta har... (More)
Popular Abstract in Swedish

I cellen styrs allt av biomolekylernas förmåga till igenkänning och växelverkan med varandra. Molekylernas speciella egenskaper bygger upp kroppens alla komplicerade funktioner, till exempel immunförsvar, celldelning och muskelarbete. Interaktionerna mellan biomolekylerna kan vara av olika karaktär; de beskrivs som starka eller svaga (affinitet), långsamma eller snabba (kinetik).



Det har visat sig att många biologiska mekanismer styrs av svaga interaktioner som, flera tillsammans, skapar igenkänning och stringens. Tack vare att de enskilda interaktionerna är svaga skapas samtidigt en dynamik och förmåga för systemen att snabbt anpassa sig till omgivande förhållanden. Detta har samtidigt medfört att de varit har svåra att studera, just på grund av att de är så svaga.



Denna avhandling handlar om dessa svaga biomolekylära interaktioner. Den diskuterar vilken roll de har i biokemiska kommunikationer och hur man kan studera dessa. Tonvikten ligger dock på hur man kan utnyttja svagt växelverkande biomolekyler som reagens i analytiska metoder. Som reagens använder man också antikroppar. Dessa proteiner är en viktig del i kroppens immunförsvar och kan på konstgjord väg fås att känna igen de speciella molekyler (antigen) man är intresserad av. Antikropparna används sedan i analyser av exempelvis läkemedel, diagnostik eller i biotekniska processer.



I de arbeten som presenteras här har låg-affina antikroppar använts i olika modellsystem.



Artikel I visar att svaga interaktioner lämpar sig väl för kromatografisk separation av strukturellt närbesläktade molekyler. Övriga arbeten handlar om hur svaga interaktioner kan tillämpas i biosensorer.



I artiklarna II och III utforskas möjligheterna till mätning av verkligt svaga biomolekylinteraktioner. Den dynamik och specificitet som kännetecknar dessa gör att man kan använda dem till att övervaka föränderliga processer.



Artiklarna IV och V introducerar den kontinuerliga immunsensorn som har stora möjligheter till att användas i områden där man vill mäta fluktuerande koncentrationer av intressanta molekyler. (Less)
Abstract
Molecular recognition and interaction is fundamental for the function of biological systems. The properties of the interacting biomolecules dictate the type of forces involved and the strength and dynamics of the interaction. Many interactions are very strong whereas others exhibit weak affinity.



Typically, weak interactions work in concert to trigger a biological response. The advantage with this approach is the inherent dynamics. It has been shown that this approach can be successful for in vitro applications as well. By exposing analytes to a multitude of specific, weak affinity interactions, which are governed by fast association and dissociation rates, separation based on small differences in affinity is possible.... (More)
Molecular recognition and interaction is fundamental for the function of biological systems. The properties of the interacting biomolecules dictate the type of forces involved and the strength and dynamics of the interaction. Many interactions are very strong whereas others exhibit weak affinity.



Typically, weak interactions work in concert to trigger a biological response. The advantage with this approach is the inherent dynamics. It has been shown that this approach can be successful for in vitro applications as well. By exposing analytes to a multitude of specific, weak affinity interactions, which are governed by fast association and dissociation rates, separation based on small differences in affinity is possible. The same principles can also be used to characterize biological weak affinity binders and for analytical purposes.



This thesis has discussed biomolecular interactions in the weak affinity range (defined in this investigation as dissociation constants (KD) larger than 0.01 mM) in general and has focuses on how they can be studied and exploited in vitro. Weak affinity monoclonal antibodies were used as model systems in three different applications:



(i) to explore how weak affinity chromatography based on monoclonal IgM can be employed to separate structurally related steroids under non-denaturing conditions.



(ii) to investigate the possibilities of studying weak interactions between antibodies and haptens with a real-time optical biosensor based on surface plasmon resonance. A rationale for the design of such experiments to avoid inaccurate results was suggested



(iii) to introduce continuous real-time immunosensing for monitoring fluctuating concentrations of biomolecules in a flow. (Less)
Please use this url to cite or link to this publication:
author
opponent
  • Prof. van Regenmortel, Marc, CNRS, Strasbourg, France
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Bioteknik, Biotechnology, immunosensor, biosensor, weak affinity chromatography, cross-reactivity, antibody-antigen interaction, weak affinity, biomolecular recognition, Immunology, serology, transplantation, Immunologi, serologi
pages
98 pages
publisher
Magnus Strandh, Department of Chemistry and Biomedical Sciences, University of Kalmar, SE-391 82 Kalmar, Sweden,
defense location
Hörsalen Falken, Nygatan 18b, Kalmar
defense date
2000-11-17 10:30
external identifiers
  • Other:ISRN: LUTKDH/TKBK-00/1002--SE
ISBN
91-628-4481-4
language
English
LU publication?
yes
id
130e2cdf-8680-46d8-a0f2-4ad97ff2e366 (old id 40989)
date added to LUP
2007-08-01 12:21:14
date last changed
2016-09-19 08:45:14
@misc{130e2cdf-8680-46d8-a0f2-4ad97ff2e366,
  abstract     = {Molecular recognition and interaction is fundamental for the function of biological systems. The properties of the interacting biomolecules dictate the type of forces involved and the strength and dynamics of the interaction. Many interactions are very strong whereas others exhibit weak affinity.<br/><br>
<br/><br>
Typically, weak interactions work in concert to trigger a biological response. The advantage with this approach is the inherent dynamics. It has been shown that this approach can be successful for in vitro applications as well. By exposing analytes to a multitude of specific, weak affinity interactions, which are governed by fast association and dissociation rates, separation based on small differences in affinity is possible. The same principles can also be used to characterize biological weak affinity binders and for analytical purposes.<br/><br>
<br/><br>
This thesis has discussed biomolecular interactions in the weak affinity range (defined in this investigation as dissociation constants (KD) larger than 0.01 mM) in general and has focuses on how they can be studied and exploited in vitro. Weak affinity monoclonal antibodies were used as model systems in three different applications:<br/><br>
<br/><br>
(i) to explore how weak affinity chromatography based on monoclonal IgM can be employed to separate structurally related steroids under non-denaturing conditions.<br/><br>
<br/><br>
(ii) to investigate the possibilities of studying weak interactions between antibodies and haptens with a real-time optical biosensor based on surface plasmon resonance. A rationale for the design of such experiments to avoid inaccurate results was suggested<br/><br>
<br/><br>
(iii) to introduce continuous real-time immunosensing for monitoring fluctuating concentrations of biomolecules in a flow.},
  author       = {Strandh, Magnus},
  isbn         = {91-628-4481-4},
  keyword      = {Bioteknik,Biotechnology,immunosensor,biosensor,weak affinity chromatography,cross-reactivity,antibody-antigen interaction,weak affinity,biomolecular recognition,Immunology,serology,transplantation,Immunologi,serologi},
  language     = {eng},
  pages        = {98},
  publisher    = {ARRAY(0xabbf408)},
  title        = {Insights into weak affinity antibody-antigen interactionsStudies using affinity chromatography and optical biosensor},
  year         = {2000},
}