Determination of dissociation constants by affinity electrophoresis: Complexes between human serum proteins and concanavalin A

Bøg-Hansen, T.C.; Takeo, Kazusuke

doi:10.1002/elps.1150010112

Cited by 72 publications

(13 citation statements)

References 22 publications

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“…This would also be supported by considering the large difference in the concentration of LCA needed to produce half maximum inhibition and activation of the MoAb binding to AFP-L3. The LCA concentration needed to produce half maximum inhibition was close to the dissociation constant of AFP-L3-LCA complex (K d , 0.067 mg/ml at 5°C and 0.24 mg/ml at 25°C) determined by lectin affinity electrophoresis [7] and applying the theory of Bøg-Hansen and Takeo [8]. However, the concentrations of LCA needed for half-maximum activation were much less and a high concentration of LCA rather reduced the extent of enhanced binding.…”

Section: Discussionmentioning

confidence: 99%

Lectin-Dependent Modulation of Interaction between Human Alpha-Fetoprotein and Its Monoclonal Antibodies

Taketa¹,

Kamakura²,

Satomura³

et al. 1998

Tumor Biol

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The effect of lentil lectin (LCA) on the binding of mouse monoclonal antibody (MoAb) against human α-fetoprotein (AFP) to LCA-nonreactive AFP-L1 and LCA-reactive AFP-L3 was studied on a panel of 30 MoAbs provided by the TD-2 Workshop of ISOBM for epitope mapping. LCA inhibited the binding of MoAbs 93 and 98 to AFP-L3 but not to AFP-L1, indicating that there was a competition between the MoAbs and LCA for the AFP sugar chain. With MoAbs 100, 109, 118, and 120, LCA rather increased the binding to AFP-L3 over that to AFP-L1. These modulating effects of LCA on the MoAb binding to AFP-L3 were abolished by periodate treatment of the AFP preparations without affecting the binding of MoAb to AFP. Concanavalin A had similar inhibiting and enhancing effects on MoAb binding, but equally to AFP-L1 and AFP-L3, both of which are fully reactive with concanavalin A. The results suggested that MoAbs 93 and 98 recognized epitopes closely related to sugar chain, and their binding to AFP-L3 was inhibited by the bound LCA due to steric hindrance. The enhanced binding of some MoAbs to AFP-L3 over AFP-L1 with LCA, or both glycoforms of AFP with concanavalin A, may be explained by postulating an allosteric mechanism mediated by the oligosaccharide-lectin interaction.

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Section: Discussionmentioning

confidence: 99%

Lectin-Dependent Modulation of Interaction between Human Alpha-Fetoprotein and Its Monoclonal Antibodies

Taketa¹,

Kamakura²,

Satomura³

et al. 1998

Tumor Biol

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“…Dissociation constants of AFP-E,-PHA complexes were calculated from the relative mobilities of AFP glycoforms at different concentrations of E,-PHA by the BsgHansen and Takeo plot [8] as shown in Fig.3. Dissociation constants and relative mobilities of AFP-E,-PHA complexes (RmJ calculated from Fig.…”

Section: Resultsmentioning

confidence: 99%

Lectin‐gradient gel affinity electrophoresis of glycoproteins

et al. 1998

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Lectin-gradient agarose gel affinity electrophoresis was developed for identification of glycoforms of glycoproteins in lectin affinity electrophoresis. Gradation of lectin was done by stacking agarose gel blocks with increasing concentrations of lectin (discontinuous system) and by keeping the plate in a moist chamber at 4 degrees C overnight (continuous system) before electrophoresis. On the visualization of separated glycoform lines, the antibody-affinity blotting was superior for low concentrations of alpha-fetoprotein. Fluoresceine isothiocyanate (FITC) labeling of whole serum proteins, enzyme activity staining for alkaline phosphatase, and prestaining for lipoproteins were also applicable for visualization of proteins at higher concentrations. The conventional Western blotting can not be recommended because of the competition between lectin and glycoproteins in binding to nitrocellulose membrane. Lectin-gradient affinity electrophoresis also had a wide application for optimization of the condition of lectin affinity electrophoresis.

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“…Kinetic and electrophoretic constants of AFP bands separated with DSA were deter mined according to the formula of BogHansen and Takeo [24] and are given in table 1. AFP-D4 had the highest affinity among the AFP bands separated with DSA with an apparent dissociation constant (K<j) of 0.25 X 10-6 M. When the K«, of AFP-D4 was compared with those of AFP bands sep arated with other lectins, AFP-D4 had the lowest K<j, which was nearly one tenth the next lowest of 2.2-2.5 X 10~6MforAFP-L3 [23].…”

Section: Resultsmentioning

confidence: 99%

<i>Datura stramonium</i> Agglutinin-Reactive α-Fetoprotein Isoforms in Hepatocellular Carcinoma and Other Tumors

Taketa¹,

Ichikawa²,

Yamamoto³

et al. 1990

Tumor Biol

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By means of Datura stramonium agglutinin (DSA) affinity electrophoresis, human α-fetoprotein (AFP) was resolved into five bands, AFP-D1, D2, D3, D4 and D5, in order of decreasing mobility. AFP-D1, which had no affinity for DSA, comprised more than 84% of the intensity of total AFP bands. The percentage of AFP-D2 increased marginally in hepatitis and liver cirrhosis with or without hepatocellular carcinoma. AFP-D3 increased characteristically in hepatocellular carcinoma and AFP-D4, which had the highest affinity for DSA, increased up to 12% in other tumors, mostly of gastrointestinal origin. AFP-D5 showed no consistent changes among the benign and malignant diseases. The assay of AFP-D3 and D4 proved useful as a highly specific marker of hapatocellular carcinoma and other tumors, respectively.

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Determination of dissociation constants by affinity electrophoresis: Complexes between human serum proteins and concanavalin A

Abstract: Affinity electrophoresis Electrophoresis 1980, I , 67-71 electrofocusing in that the former revealed greater heterogeneity of components detected.

Cited by 72 publications

References 22 publications

Lectin-Dependent Modulation of Interaction between Human Alpha-Fetoprotein and Its Monoclonal Antibodies

Lectin-Dependent Modulation of Interaction between Human Alpha-Fetoprotein and Its Monoclonal Antibodies

Lectin‐gradient gel affinity electrophoresis of glycoproteins

<i>Datura stramonium</i> Agglutinin-Reactive α-Fetoprotein Isoforms in Hepatocellular Carcinoma and Other Tumors

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Determination of dissociation constants by affinity electrophoresis: Complexes between human serum proteins and concanavalin A

Abstract: Affinity electrophoresis Electrophoresis 1980, I , 67-71 electrofocusing in that the former revealed greater heterogeneity of components detected.

Cited by 72 publications

References 22 publications

Lectin-Dependent Modulation of Interaction between Human Alpha-Fetoprotein and Its Monoclonal Antibodies

Lectin-Dependent Modulation of Interaction between Human Alpha-Fetoprotein and Its Monoclonal Antibodies

Lectin‐gradient gel affinity electrophoresis of glycoproteins

<i>Datura stramonium</i> Agglutinin-Reactive &alpha;-Fetoprotein Isoforms in Hepatocellular Carcinoma and Other Tumors

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<i>Datura stramonium</i> Agglutinin-Reactive α-Fetoprotein Isoforms in Hepatocellular Carcinoma and Other Tumors