Detection and characterization of weak affinity antibody antigen recognition with biomolecular interaction analysis

Ohlson, Sten; Strandh, Magnus; Nilshans, Helena

doi:10.1002/(sici)1099-1352(199705/06)10:3<135::aid-jmr355>3.0.co;2-b

Cited by 41 publications

(17 citation statements)

References 8 publications

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“…[29] Moreover, to obtain sufficient SPR signal for the weak-affinity binding of a carbohydrate to a protein with analytes of molecular mass < 1000 Da, high analyte concentrations (up to the millimolar range) are required. [15,[30][31][32][33][34] Under these conditions, the contribution from the bulk refractive index to the specific response becomes significant, with an apparent loss in specific binding.…”

Section: Introductionmentioning

confidence: 93%

SPR Studies of Carbohydrate–Protein Interactions: Signal Enhancement of Low‐Molecular‐Mass Analytes by Organoplatinum(II)‐Labeling

et al. 2005

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The relatively insensitive surface plasmon resonance (SPR) signal detection of low‐molecular‐mass analytes that bind with weak affinity to a protein—for example, carbohydrate–lectin binding—is hampering the use of biosensors in interaction studies. In this investigation, low‐molecular‐mass carbohydrates have been labeled with an organoplatinum(II) complex of the type [PtCl(NCNR)]. The attachment of this complex increased the SPR response tremendously and allowed the detection of binding events between monosaccharides and lectins at very low analyte concentrations. The platinum atom inside the organoplatinum(II) complex was shown to be essential for the SPR‐signal enhancement. The organoplatinum(II) complex did not influence the specificity of the biological interaction, but both the signal enhancement and the different binding character of labeled compounds when compared with unlabeled ones makes the method unsuitable for the direct calculation of biologically relevant kinetic parameters. However, the labeling procedure is expected to be of high relevance for qualitative binding studies and relative affinity ranking of small molecules (not restricted only to carbohydrates) to receptors, a process of immense interest in pharmaceutical research.

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Section: Introductionmentioning

confidence: 93%

SPR Studies of Carbohydrate–Protein Interactions: Signal Enhancement of Low‐Molecular‐Mass Analytes by Organoplatinum(II)‐Labeling

et al. 2005

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“…An important feature of SPR is the possibility to obtain realtime data on the interaction of a ligand to its receptor allowing kinetic data to be determined (Day et al, 2002;Myszka et al, 2003;Ohlson et al, 1997). Moreover, the amount of material necessary to perform the experiments is small, labelling is not required and variation in surface chemistries is possible, allowing various immobilisation strategies and interaction experiments.…”

Section: Introductionmentioning

confidence: 65%

Development of an on-line SPR-digestion-nanoLC-MS/MS system for the quantification and identification of interferon-γ in plasma

Stigter

Jong

Bennekom

2009

Biosensors and Bioelectronics

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“…When soluble fibrin was injected over a surface with immobilized myosin (Figure 6B), the SPR response produced the square pulse signals typical of weak ligand interaction. 36,37 It is noteworthy that when in the latter BIA setting the analyte was prevented in its aggregation, the interaction occurred in the time scale of seconds, strengthening the interpretation of the plasmon resonance curves in Figure 6A as fibrin-initiated aggregation. The equilibrium dissociation constant calculated for the interaction of this soluble fibrin species and myosin is 0.94 M (Figure 6B inset).…”

Section: Resultsmentioning

confidence: 99%

“…36,37 The acceleration in the interaction kinetics again raises the possibility for interference of myosin with the formation of the fibrin gel. Despite the accelerated association rate of myosin and fibrin instead of fibrinogen, however, the higher affinity of the selfaggregating fibrin monomers (K d ϭ 0.156 M) 43 dominates and apparently the fibrin architecture is not affected as previously mentioned.…”

Section: Discussionmentioning

confidence: 99%

Myosin: a noncovalent stabilizer of fibrin in the process of clot dissolution

Kolev

Tenekedjiev

Ajtai

et al. 2003

Blood

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Myosin modulates the fibrinolytic process as a cofactor of the tissue plasminogen activator and as a substrate of plasmin. We report now that myosin is present in arterial thrombi and it forms reversible noncovalent complexes with fibrinogen and fibrin with equilibrium dissociation constants in the micromolar range (1.70 and 0.94 M, respectively). Competition studies using a peptide inhibitor of fibrin polymerization (glycl-prolyl-arginyl-proline [GPRP]) indicate that myosin interacts with domains common in fibrinogen and fibrin and this interaction is independent of the GPRP-binding polymerization site in the fibrinogen molecule. An association rate constant of 1.81 ؋ 10 2 M ؊1 ⅐ s ؊1 and a dissociation rate constant of 3.07 ؋ 10 ؊4 s ؊1 are determined for the fibrinogen-myosin interaction. Surface plasmon resonance studies indicate that fibrin serves as a matrix core for myosin aggregation. The fibrin clots equilibrated with myosin are stabilized against dissolution initiated by plasminogen and tissuetype plasminogen activator (tPA) or urokinase (at fibrin monomer-myosin molar ratio as high as 30) and by plasmin under static and flow conditions (at fibrin monomer-myosin molar ratio lower than 15).Myosin exerts similar effects on the tPAinduced dissolution of blood plasma clots. Covalent modification involving factor XIIIa does not contribute to this stabilizing effect; myosin is not covalently attached to the clot by the time of complete cross-linking of fibrin. Thus, our in vitro data suggest that myosin detected in arterial thrombi binds to the polymerized fibrin, in the bound form its tPA-cofactor properties are masked, and the myosin- IntroductionThe appropriate timing and localization of the proteolytic removal of the fibrin clots in the vascular bed is based on elaborate regulatory mechanisms that determine the availability of fibrinolytic proteases at the level of plasminogen activation 1 and inactivation of the enzymes. 2 Quantitative 3,4 and morphologic 5 data have revealed the importance of the structure of the fibrin substrate in determining the efficiency of fibrinolysis; the fiber diameter, the pore size of the gel, and the frequency of branching points profoundly affect the rate of fibrin dissolution. In experimental models it is rather convenient to modify the structural characteristics of the fibrin gel by changing the concentrations of thrombin and fibrinogen when fibrin is generated 6,7 or the ionic strength at polymerization 4,8 for evaluation of the efficiency of fibrinolysis on various fibrin substrates. In vivo, however, fibrinogen, the precursor of fibrin, circulates in blood surrounded by cells and proteins. Thus, when thrombin converts it to fibrin, the polymerization occurs in a milieu highly enriched in macromolecules. The fibrin assembly is definitely modified by the presence of plasma proteins (eg, albumin 9 ), cellular elements (eg, erythrocytes 10 ), or specific proteins of isolated compartments (eg, amyloid ␤-protein 11 ). This modification can be attributed to the effect of vo...

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Detection and characterization of weak affinity antibody antigen recognition with biomolecular interaction analysis

Cited by 41 publications

References 8 publications

SPR Studies of Carbohydrate–Protein Interactions: Signal Enhancement of Low‐Molecular‐Mass Analytes by Organoplatinum(II)‐Labeling

SPR Studies of Carbohydrate–Protein Interactions: Signal Enhancement of Low‐Molecular‐Mass Analytes by Organoplatinum(II)‐Labeling

Development of an on-line SPR-digestion-nanoLC-MS/MS system for the quantification and identification of interferon-γ in plasma

Myosin: a noncovalent stabilizer of fibrin in the process of clot dissolution

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