Analysis of macromolecular interactions using immobilized ligands

Chaiken, Irwin M.; Rosé, Sergio D.; Karlsson, Robert

doi:10.1016/0003-2697(92)90329-6

Cited by 194 publications

(129 citation statements)

References 35 publications

Supporting

Mentioning

125

Contrasting

Order By: Relevance

“…12,13 However, it has been shown that the k d values for the surface-immobilized probe-ligand systems are typically several orders of magnitude larger, and values as high as 10 À7 have been recorded for immobilized biotin-streptavidin systems. 11,14 This significant difference in binding behavior suggests that steric hindrance plays a role in the interaction of the binding pair when one half is immobilized. 9,11,15,16 Additionally, prior literature shows that the dissociation constant for this binding pair can vary significantly depending on the specific environment and test conditions during association and dissociation phases, as expected.…”

Section: à14mentioning

confidence: 99%

“…By adding a transport constant and performing experiments over a wide range of flow rates and concentrations, it would possible to correct for these effects and to accurately determine the k a . 14,16 Additionally, as the majority of previous experiments with microspheres have been performed in non-flowing or stagnant solutions, optimizing the flow rate could provide a route to improve the collection efficiency or the detection ability of microsphere sensors. Results of flow rate optimization experiments would have a wide range of applications including, but not limited to, the arena of binding kinetics.…”

Section: à14mentioning

confidence: 99%

See 1 more Smart Citation

Determination of binding kinetics using whispering gallery mode microcavities

Soteropulos

Hunt

Armani

2011

Applied Physics Letters

View full text Add to dashboard Cite

Silica optical microcavity sensors show great promise in the kinetic evaluation of binding pairs, fundamental in understanding biomolecular interactions. Here, we develop and demonstrate a novel platform, based on bioconjugated silica microsphere resonators, to study the binding kinetics of the biotin-streptavidin system. We characterize the optical performance, verify the covalent attachment of biotin to the surface, and perform streptavidin detection experiments. We perform preliminary kinetic analysis of the detection data which shows the potential of whispering gallery mode resonators in the determination of the dissociation constant of the binding pair, which is in good agreement with previously published values. The binding site affinity of an enzyme or antibody (i.e., its ability to bind to its specific complement), determines its utility in a given application. For example, in designing therapeutics, a strong affinity for a specific target is desired to maximize the effectiveness of the therapeutic.1 Conversely, in immunoassay applications, a tunable affinity is often desired to enable sensor reusability.2 Therefore, it is not surprising that the study of binding site affinities is of great scientific and industrial importance. In order to fully characterize the binding site affinity, it is necessary to perform measurements using both free and fixed molecules. However, while there are numerous methods for determining the affinity with free molecules, the approaches for fixed molecules are limited. 2The primary approach for fixed molecule characterization is based on surface plasmon resonance (SPR) sensors. This technique relies on the decaying evanescent optical field generated by the SPR, which is established at the interface of a metal film and the solution, and is approximately $50 nm long.3 When a molecule is located within the evanescent field, it modifies the effective refractive index. This change is detected as an increase in the SPR wavelength. While current SPR systems are ideal for characterizing single interaction pairs, they often experience difficulties resolving multi-valent interactions. This limitation is a result of the short interaction time between the photon and the molecule, limited bioconjugation protocols for attachment of one half of the binding pair to metal surfaces, and restricted mass transport across the sensor surface.

show abstract

Section: à14mentioning

confidence: 99%

Section: à14mentioning

confidence: 99%

Determination of binding kinetics using whispering gallery mode microcavities

Soteropulos

Hunt

Armani

2011

Applied Physics Letters

View full text Add to dashboard Cite

show abstract

“…The BIAcore (biospecific interaction analysis) system, developed recently by Pharmacia Biosensor to measure surface plasmon resonance (SPR), allows interactions between two macromolecules to be monitored in real time [21,22]. This system has been used for analysis of interactions between *Corresponding author.…”

Section: Introductionmentioning

confidence: 99%

Kinetics of binding of Antp homeodomain to DNA analyzed by measurements of surface plasmon resonance

Seimiya¹,

Kurosawa²

1996

FEBS Letters

View full text Add to dashboard Cite

The kinetics of binding of the Antp homeodomain to three kinds of DNA fragment were analyzed by measurements of surface plasmon resonance at various temperatures. Non-specific and specific binding of the homeodomain to DNA was examined. In the case of non-specific binding, the association rate constant (kass) was estimated to be 1.41-2.62X 105 M -1 s -1 and the dissociation rate constant (kdiss) was 1.36-3.10 X 10 -2 s -1, thus, the dissociation constant (KI)) was 0.847-1.72 × 10 7 M. The association seemed to be driven by entropy. In the case of specific binding, by contrast, the enthalpy term seemed to contribute more to the binding than did the entropy term. The kass was 2.04-2.59X105 M -1 s -1 and the kdiss was 0.759-1.16X10 -3 s -1, thus, the /to was 2.93-5.69X 10 -9 M. These values were measured under the condition of 150 mM NaCl. Both interactions were strongly dependent on the concentration of NaC1. The /to at 50 mM NaCI became several tens of times smaller than those at 150 mM. Possible reasons for the differences between non-specific and specific interactions are discussed.

show abstract

“…2) (M.-L.L., K. Alfthan and K.K., manuscript in preparation). In the latter method, association and dissociation of sample molecules in a layer of dextran-immobilized ligand give rise to changes in the refractive index, which is monitored in real time [26]. Immunoliposomes display markedly slower dissociation from the immobilized hapten than the soluble antibody due to multivalent binding as there are many antibody molecules per liposome (Fig.…”

Section: Genetically Lipid-modified Antibodiesmentioning

confidence: 99%