Determination of antibody affinity and kinetic binding constants in Gyrolab Bioaffy microfluidic CD
Independent thesis Basic level (professional degree), 20 points / 30 hpStudent thesis
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.
Affinity constants, Equilibrium dissociation constants, Kinetic rate constants, Immunoassay, Monoclonal antibodies, Microfluidics, Gyrolab Bioaffy, Microlaboratory, Compact disc
IdentifiersURN: urn:nbn:se:liu:diva-11616ISRN: LITH-IFM-A-EX-08/1931-SEOAI: oai:DiVA.org:liu-11616DiVA: diva2:18035
2008-04-01, Planck, Fysikhuset, IFM / Linköpings Universitet, Linköping, 15:15
Hoffmann, StephanEngström, Johan