The influence of adsorption of native and modified antibodies on their activity

Lin, Jinn-Nan; Andrade, Joseph D.; Chang, I‐Nan

doi:10.1016/0022-1759(89)90079-3

Cited by 52 publications

(34 citation statements)

References 15 publications

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“…Overall, over the low antibody surface excess range, the binding ratio of hCG to antibody could be as high as 0.7, equivalent to 0.35 hCG binding to each Fab site at the interface. While the surface activity of the immobilized antibody is indeed broadly similar to that reported in the literature, [11][12]22,[49][50][51] this study demonstrates a quantitative variation of antibody binding activity with its surface excess…”

Section: Kinetic and Equilibrated Adsorptionsupporting

confidence: 72%

Effect of Surface Packing Density of Interfacially Adsorbed Monoclonal Antibody on the Binding of Hormonal Antigen Human Chorionic Gonadotrophin

2006

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Interfacial adsorption of a mouse monoclonal antibody (type IgG1, anti-beta-hCG) at the hydrophilic silicon oxide/water interface has been studied by spectroscopic ellipsometry and neutron reflection, followed by assessment of binding of a hormonal antigen, human chorionic gonadotrophin (hCG), onto the adsorbed antibody molecules. The amount of adsorption reached a maximum around the isoelectric pH (IP) of 6 for the antibody; this pH-dependent pattern could be altered by increasing salt concentration, a trend also observed for other proteins. Neutron reflection revealed the formation of a 40 A uniform layer from the adsorbed antibody, indicating a flat-on orientation. The subsequent hCG binding showed that the molar ratio of hCG bound to antibody at the interface was as high as 0.7 at low surface coverage of antibody and decreased with increasing surface antibody concentration. The results point to an increasing extent of steric hindrance to hCG access with increasing packing density of antibody molecules on the surface. Comparison with previously published crystal structure studies suggests twisting of the variable region to allow access of the antigen. The binding of hCG was also found to be pH-dependent with its maximum around the IP, if the ionic strength of the solution was low (20 mM). However, if the ionic strength was increased to 200 mM, then hCG binding was influenced by a combination of steric hindrance and electrostatic interaction between the antigen and the surface. These results are highly relevant to the improvement of the performance of biotechnologies such as fertility test pads and biosensors based on antibody immobilization.

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Section: Kinetic and Equilibrated Adsorptionsupporting

confidence: 72%

Effect of Surface Packing Density of Interfacially Adsorbed Monoclonal Antibody on the Binding of Hormonal Antigen Human Chorionic Gonadotrophin

2006

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“…Radiolabeling the protein with 125 I before immobilization was used to quantify the average surface coverage. 13 The fluorescein titration of the specific anti-fluorescyl IgG silica surfaces was carried out using total internal reflection fluorescence (TIRF) spectroscopy 14 in order to establish the functionality and the binding affinity of the immobilized anti-fluorescyl IgG.…”

Section: Methodsmentioning

confidence: 99%

Effects of Discrete Protein-Surface Interactions in Scanning Force Microscopy Adhesion Force Measurements

Stuart

Hlady

1995

Langmuir

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The potential for measuring specific molecular recognition forces between probe-bound ligands and surface-bound proteins using a scanning force microscope (SFM) has recently gained much attention. Generally, observed discontinuities in the SFM force-displacement curves are attributed to the breaking of discrete, specific affinity bonds. The present study on the molecular recognition system composed of surface-immobilized antifluorescyl IgG molecules and SFM probe-bound fluorescein ligands has demonstrated that similar intermittent discontinuities in the SFM forcedisplacement curves may in fact be largely due to nonspecific discrete interactions between the protein and the SFM probe. The mechanical behavior of the cantilever-spherical bead system used in this study is discussed, as it appears to cause a false indication of the separation distance between the surface and probe. The strong lateral interactions which result in "stick and slip"-like discontinuities seen in the adhesion curves are likely the result of localized adhesion due to the heterogeneous nature of proteins and the lack of molecular mobility allowed in the experimental system. The effect is magnified with increasing contact time between the protein and probe. Factors which may cause such anomalous behavior in a specific ligand-protein system are discussed in order to avoid misinterpretation of SFM adhesion measurements.

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“…In this system, the solid support is also the antigen, and so part of these sites are occupied by the blocking agent. Furthermore, as antibodies may easily denature after adsorption on a solid surface (27,28), their adsorption on the crystal may be partially or totally irreversible. Thus, the equilibrium constant for adsorption on crystals is not amenable to be expressed as a function of a known concentration of bound, relative to free, antibody.…”

Section: The Resultsmentioning

confidence: 99%

Monoclonal antibodies that specifically recognize crystals of dinitrobenzene

Kessler

Perl-Treves²,

Addadi³

1996

FASEB j.

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Monoclonal antibodies have been elicited and selected after injection of crystals of 1,4-dinitrobenzene (1,4-DNB) and cholesterol monohydrate in mice. The reactivity of some of these antibodies to 1,4-DNB crystals, cholesterol monohydrate crystals, and other solid substrates has been characterized. Two of the antibodies selectively recognize 1,4-DNB crystal surfaces in an appropriately modified ELISA. They do not interact either with 1,4-DNB/BSA conjugates or with polystyrene and cholesterol monohydrate surfaces. They do interact with 1,2-DNB crystal surfaces, albeit with much lower reactivity. It is consequently suggested that these antibodies are not specific to the DNB molecule but rather to a repetitive motif of molecular moieties exposed at the crystal surfaces. Characterization of their binding regions may help to elucidate the interactions of antibodies with solid substrates and, in general, with antigens exposed on biological and artificial surfaces.

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The influence of adsorption of native and modified antibodies on their activity

Cited by 52 publications

References 15 publications

Effect of Surface Packing Density of Interfacially Adsorbed Monoclonal Antibody on the Binding of Hormonal Antigen Human Chorionic Gonadotrophin

Effect of Surface Packing Density of Interfacially Adsorbed Monoclonal Antibody on the Binding of Hormonal Antigen Human Chorionic Gonadotrophin

Effects of Discrete Protein-Surface Interactions in Scanning Force Microscopy Adhesion Force Measurements

Monoclonal antibodies that specifically recognize crystals of dinitrobenzene

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