Investigation of the antigen-antibody rection by fluorescence polarization

Kierszenbaum, Felipe; Dandliker, J; Dandliker, Walter B.

doi:10.1016/0019-2791(69)90184-0

Cited by 23 publications

(7 citation statements)

References 21 publications

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“…(3), the intrinsic binding constant of the monoclonal mouse anti-BSA is then 3.5 * 0.6 X lo7 M-'. This result is about 10 times lower than that previously reported for a rabbit polyclonal anti-BSA (3.7 X 10' M-l) using a fluorescence polarization method [38]. Labeling is an important factor in the difference, since those authors also reported a 40% decrease in binding constant for anti-BSA when labeled antigen (2.7 dansyl residues per BSA*) was used instead of unlabeled antigen.…”

Section: Affinity Constants and Complex Speciationcontrasting

confidence: 70%

Monoclonal antibody binding affinity determined by microchip‐based capillary electrophoresis

1998

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The affinity constant of a monoclonal antibody to fluorescently labeled bovine serum albumin (BSA) was measured in diluted mouse ascites fluid using a microfluidic chip to perform affinity capillary electrophoresis. Borofloat glass-based devices could be used repeatedly with samples for many months. On-chip separations were performed in less than 60 s, and 30-60 s was required for manual sample exchange. The change in peak height for BSA with increasing BSA/anti-BSA concentration ratio was used to determine concentration changes in bound and free BSA. A Scatchard plot analysis gave an affinity constant (more exactly the intrinsic association constant) of 3.5+/-0.6 x 10(7) M(-1) for a 1:1 stoichiometric ratio. Two affinity complexes were separated. One complex was identified by the Scatchard method as having a 1:1 stoichiometric ratio. The other complex is proposed to have a stoichiometry with an excess of anti-BSA to BSA, most likely (anti-BSA)2-BSA, on the basis of a faster migration time than the 1:1 complex, a decrease in the amount of this complex with increasing [BSA], and predictions of theoretical models for multi-valent antigens. Potential applications of microchip-based devices in affinity measurements are discussed.

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Section: Affinity Constants and Complex Speciationcontrasting

confidence: 70%

Monoclonal antibody binding affinity determined by microchip‐based capillary electrophoresis

1998

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“…Application of simplex optimization for maximizing the F ratio of multiple-regression analysis to fit equilibrium sedimentation patterns with a constraint for rejecting negative weight fractions yielded accurate estimates for molecular weight and composition of the components in mixtures of up to six components, if selection of sample " Green (1963).6 Kuzmanoff et al (1990).1 : Kierszenbaum et al (1969). d Parham (1984).…”

Section: Discussionmentioning

confidence: 99%

Computation of molecular weight and weight fraction of five and six components in mixtures from model equilibrium ultracentrifugation data

Nakai¹,

Nonaka²

1992

J. Agric. Food Chem.

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“…Fluorescence polarization of a macromolecule is mainly dependent on the geometry and rigidity of the molecular structure. Changes in molecular conformation due to aggregation and interaction will, therefore, result in changes in the polarization, because these changes invite a modification of molecular geometry and volume in the macromolecule (Weber, 1952;Steiner, 1954;Haber and Bennett, 1962;Kierszenbaum et al,, 1969).…”

Section: Theorymentioning

confidence: 99%

“…. (1964) andKierszenbaum et al (1969) reported a method for estimation of the association constant between antigen and antibody by fluorescence polarization. The basic equation in those papers and ours are the same.…”

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confidence: 99%

Investigation of κ-αS1-casein interaction by fluorescence polarization

Clarke¹,

Nakai²

1971

Biochemistry

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The interaction between aSl-and K-casein was studied by the fluorescence polarization technique. The entropy and enthalpy changes, AS" and AH", were positive suggesting that the ease or spontaneity of interaction increases with temperature, and that the interaction is probably hydrophobic in nature. The results at 40 and 50" demonstrate a S ince the fractionation of a-casein into fracti0n.s as and K , calcium sensitive and insensitive, respectively (Waugh and von Hippel, 1956), there has been a great interest in the Kcasein fraction because of its unique stabilizing ability toward the other caseins.Studies have been done on the interaction between K -and aI-caseins, which stabilizes a,l-casein against calcium precipitation, and could act as precursor to stable casein micelles Waugh and Noble, 1965).The techniques used and the results obtained were varied regarding the interacting ratio of a r~-to K-casein and the type of interaction, i.e., hydrophilic, hydrophobic, or electrostatic. Waugh and von Hippel (1956) studied the interaction between cyE1-and K-casein and found a variable interaction ratio, but found that the cySl : K ratio was predominantly 4. found that the aSl : K ratio was low and close t o unity. Garnier (1967) found an interaction ratic of 1 mole of aS1-to 3 moles of K-casein. Howeker, in an earlier paper, Garnier et ai. (1964a) reported a n interaction ratio of unity. Parry et al. (1969) found the interaction ratio to be unity, using gel filtration. Kenkare and Hansen (1967) found that the interaction product was of variable composition, and suggested that the interaction between the cyysl-and K-casein was hydrophilic in nature. This is in contrast with the work of Garnier et al. (1964b) who found a positive entropy for the reaction, suggestive of hydrophobic interaction.This postulation seems to be feasible as the interaction was inhibited at 2-6" and spontaneous above 37" .Payens (1 966) suggested that the main interacting force between the caseins, in maintaining the micelle structure, is hydrophobic in nature within the micelle. This concept was based on the amino acid composition of the caseins, the temperature dependence of aggregation of the caseins and the established hydrophobic nature of aggregation of cysland /3-caseins.Hill and Wake (1969) discussed the amphiphilic nature of K-casein and suggested that the cyal-n-casein interaction is significantly hydrophobic in nature.

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Investigation of the antigen-antibody rection by fluorescence polarization

Cited by 23 publications

References 21 publications

Monoclonal antibody binding affinity determined by microchip‐based capillary electrophoresis

Monoclonal antibody binding affinity determined by microchip‐based capillary electrophoresis

Computation of molecular weight and weight fraction of five and six components in mixtures from model equilibrium ultracentrifugation data

Investigation of κ-αS1-casein interaction by fluorescence polarization

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