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Determination of antibody affinity and kinetic binding constants in Gyrolab Bioaffy microfluidic CD
Linköping University, The Department of Physics, Chemistry and Biology.
2008 (English)Independent thesis Basic level (professional degree), 20 points / 30 hpStudent thesis
Abstract [en]

Studies of binding reactions are of highest importance in a vast number of areas of biomedicine and biotechnology. A demand for fast and accurate small-volume measurements grows stronger, partly due to the development of therapeutic antibodies. In this report, a novel method for studies of binding reactions of antibodies is described. The use of a microfluidic platform shows promising results in determination of affinity binding constants.

Affinities between 1E-09 and 1E-11 M have been determined for four TSH antibodies. Reproducibility tests give a CV below 10%, using different Gyrolab instruments and microfluidic CD:s. The method carries the advantages of using solution-based measurements of unmodified molecules. Also an initial proof-of-concept for measurement of binding reaction rate constants shows further usage of the method. The kinetic association rate constant has been determined to 2E+06 M-1s-1 for one antibody. The possibility of using this method for screening of antibody libraries is also discussed.

Place, publisher, year, edition, pages
Institutionen för fysik, kemi och biologi , 2008. , 59 p.
Keyword [en]
Affinity constants, Equilibrium dissociation constants, Kinetic rate constants, Immunoassay, Monoclonal antibodies, Microfluidics, Gyrolab Bioaffy, Microlaboratory, Compact disc
National Category
Industrial Biotechnology
URN: urn:nbn:se:liu:diva-11616ISRN: LITH-IFM-A-EX-08/1931-SEOAI: diva2:18035
2008-04-01, Planck, Fysikhuset, IFM / Linköpings Universitet, Linköping, 15:15
Available from: 2008-04-29 Created: 2008-04-29

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