OUTSIDE Journal Cover “Analytical Methods” December Issue

Binding affinity of biomolecular interactions can be directly extracted from measured surface plasmon resonance biosensor sensorgrams by fitting the data to the appropriate model equations. The conventional method for affinity estimation uses a series of analytes and buffers that are injected serially to a single immobilized ligand on the sensing surface, including a regeneration step between each injection, to generate information about the binding behavior. We present an alternative method to estimate the affinity using a single analyte concentration injected to multiple ligand densities in a microarray format. This parameter estimation method eliminates the need for multiple analyte injections and surface regeneration steps, which can be important for applications where there is limited analyte serum, fragile ligand-surface attachment, or the detection of multiple biomolecule interactions. The single analyte injection approach for binding affinity estimation has been demonstrated for two different interactant pairs, b2 microglobulin/anti-b2 microglobulin (b2M) and human IgG/Fab fragments of anti-human IgG (hIgG), where the ligands are printed in a microarray format. Quantitative comparisons between the estimated binding affinities measured with the conventional method are b2M: K_D=1.48±0.28 nM and hIgG: K_D=12.6±0.2 nM and for the single injection method are b2M: K_D=1.52±0.22 nM and hIgG: K_D=12.5±0.6 nM, which are in good agreement in both cases.

kinetic and affinity estimation

Kinetic and affinity estimation of the biomolecular interactant pairs using a single analyte concentration injected over multiple ligand densities in a microarray format based on surface plasmon resonance imaging.

A detailed description can be found in the full paper published in “Analytical Methods”: http://www.rsc.org/Publishing/Journals/AY/article.asp?doi=b9ay00176j

For more information please contact Ganeshram Krishnamoorthy at g.krishnamoorthy@utwente.nl

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Home News & Highlights People Vacancies Research Education & Assignments Publications Intranet Internal Links External links Movies Sitemap Search	weblink \| News archive \| Archive Highlights \| Promotie film Ana Valero OUTSIDE Journal Cover “Analytical Methods” December Issue Binding affinity of biomolecular interactions can be directly extracted from measured surface plasmon resonance biosensor sensorgrams by fitting the data to the appropriate model equations. The conventional method for affinity estimation uses a series of analytes and buffers that are injected serially to a single immobilized ligand on the sensing surface, including a regeneration step between each injection, to generate information about the binding behavior. We present an alternative method to estimate the affinity using a single analyte concentration injected to multiple ligand densities in a microarray format. This parameter estimation method eliminates the need for multiple analyte injections and surface regeneration steps, which can be important for applications where there is limited analyte serum, fragile ligand-surface attachment, or the detection of multiple biomolecule interactions. The single analyte injection approach for binding affinity estimation has been demonstrated for two different interactant pairs, b2 microglobulin/anti-b2 microglobulin (b2M) and human IgG/Fab fragments of anti-human IgG (hIgG), where the ligands are printed in a microarray format. Quantitative comparisons between the estimated binding affinities measured with the conventional method are b2M: K_D=1.48±0.28 nM and hIgG: K_D=12.6±0.2 nM and for the single injection method are b2M: K_D=1.52±0.22 nM and hIgG: K_D=12.5±0.6 nM, which are in good agreement in both cases. Kinetic and affinity estimation of the biomolecular interactant pairs using a single analyte concentration injected over multiple ligand densities in a microarray format based on surface plasmon resonance imaging. A detailed description can be found in the full paper published in “Analytical Methods”: http://www.rsc.org/Publishing/Journals/AY/article.asp?doi=b9ay00176j For more information please contact Ganeshram Krishnamoorthy at g.krishnamoorthy@utwente.nl