Crystallization and preliminary X-ray diffraction study of a chimaeric Fab′ fragment of antibody binding tumour cells

Brady, R.L.; Hubbard, Roderick E.; King, David J.; Low, D.C.; Roberts, Sally; Todd, R.

doi:10.1016/0022-2836(91)90656-q

Cited by 23 publications

(16 citation statements)

References 6 publications

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“…The baseline on the sensorgram was defined with a running buffer (10 mM HEPES + 150 mM NaCl, pH 7.4) including no analyte, and all the responses were expressed relative to this baseline level. The amounts of bound analytes (mol) were calculated from the binding at 400 s after the injection; 1,000 RU corresponds to 1 ng sizes of immunoglobulin reported by electron microscopy (EM) and X-ray crystallography are 16-19 nm for IgG and 35-39 nm for IgM, respectively (Roberts et al 1995;Perkins et al 1991;Brady et al 1991). Our line profile results show that the observed height of HIgG was similar to the reported IgG size.…”

Section: Resultssupporting

confidence: 59%

Monolayers assembled from a glycolipid biosurfactant from Pseudozyma (Candida) antarctica serve as a high-affinity ligand system for immunoglobulin G and M

et al. 2007

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A carbohydrate ligand system has been developed which is composed of self-assembled monolayers (SAMs) of mannosylerythritol lipid-A (MEL-A) from Pseudozyma antarctica, serving for human immunoglobulin G and M (HIgG and HIgM). The estimated binding constants from surface plasmon resonance (SPR) measurement were Ka = 9.4 x 10(6) M(-1) for HIgG and 5.4 x 10(6) M(-1) for HIgM, respectively. The binding site was not in the Fc region of immunoglobulin but in the Fab region. Large amounts of HIgG and HIgM bound to MEL-A SAMs were directly observed by atomic force microscopy.

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Section: Resultssupporting

confidence: 59%

Monolayers assembled from a glycolipid biosurfactant from Pseudozyma (Candida) antarctica serve as a high-affinity ligand system for immunoglobulin G and M

et al. 2007

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“…Again, a supramolecular network is observed over the substrate surface. However, within this layer globular features with diameters ranging from 15 to 17 nm can be identified, which correlates with the dimensions expected for individual IgG molecules, as estimated from X-ray crystallography [31] and electron microscopy studies [32,33]. Figure 4c displays an AFM image of an anti-ferritinfunctionalized well surface, after exposure to ferritin.…”

Section: Imaging Immunoassay Surfacessupporting

confidence: 72%

The application of force microscopy to immunodiagnostic systems: imaging and biomolecular adhesion measurements

Allen

Chen

Davies

et al. 1998

Applied Physics A: Materials Science & Processing

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Affinity chromatography separation methods, biosensing devices and many diagnostic materials rely on the immobilization of biomolecules to solid supports. For example, immunoassays employ antibody molecules immobilized so that they are able to detect antigen present in the surrounding analyte. The development of the scanning probe microscopes has provided new tools for investigating such sensor surfaces. In the first section of this paper, tapping-mode atomic force microscopy (AFM) is employed to investigate the surface topographies of functionalized immunoassay substrates. Images of each biomolecular layer in an immunoassay system are produced with molecular resolution. In the second section of the paper, we investigate the ability of AFM probes coated with ferritin molecules to map the functionality of an anti-ferritin antibody-coated surface, via biomolecular recognition processes. Novel data analysis is utilized to produce two-dimensional maps of probe-sample interaction, in which each pixel point represents a single force measurement. The experimental data obtained illustrate the potential of the AFM to be used as a tool for the investigation of both surface topography and the distribution of molecular recognition sites on sensor surfaces.Immunoassays employ antibody molecules that are typically immobilized onto solid supports so that they are able to detect antigen present in the surrounding analyte [1, 2]. The atomic force microscope (AFM) [3] possesses the ability to produce molecular resolution images of a wide range of biomolecules, including DNA [4] and individual proteins [5,6]. This has been employed to investigate the antigen binding capacities of immunoassay well surfaces [7], and to discriminate between classes of antibody and antibody fragments deposited onto the immunoassay substrate [8,9]. AFM has also more recently been employed to image the specific binding of antibody molecules to solid substrates functionalized with antigen [10]. * Corresponding author The ability of the AFM to measure directly discrete intermolecular forces of 10 pN or less was first highlighted by Hoh et al. [11]. Various groups have exploited this method to probe specific biomolecular interactions between individual receptor-ligand pairs [12][13][14][15][16][17][18][19][20][21][22][23]. Utilizing this technique we have recently directly monitored receptor-ligand interactions on the immunoassay substrate [22], and have employed ferritin-functionalized AFM probes to quantify unbinding forces between ferritin and anti-ferritin antibodies bound to a silicon surface [23].Of significant interest is the use of biomolecule functionalized AFM probes for the mapping of specific recognition sites on a sample surface. Ludwig et al. [24] have employed biotinylated AFM probes to image a sample patterned with areas of streptavidin molecules. They attributed image contrast to the specific interaction between the biotinylated probe and surface-bound streptavidin molecules. A similar approach was also adopted by Hinterdorfer et al. [21] ...

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“…A homology model for human IgG4 was constructed from crystal structures for Fab B72.3 and human IgG1 (Protein Data Bank codes 1BBJ and 1HZH) (36, 37) as described previously (20). Four sets of 5000 randomized IgG4 models were created using different hinge lengths and compositions.…”

Section: Methodsmentioning

confidence: 99%

The Fab Conformations in the Solution Structure of Human Immunoglobulin G4 (IgG4) Restrict Access to Its Fc Region

Rayner

Hui

Gor

et al. 2014

Journal of Biological Chemistry

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Background: The human IgG4 antibody subclass does not activate complement and forms half-antibodies.Results: Ultracentrifugation and x-ray/neutron scattering together with atomistic modeling revealed asymmetric concentration-dependent IgG4 solution structures.Conclusion: The complement and receptor Fc binding sites are hindered by the Fab regions, explaining loss of activity.Significance: These solution structures clarify IgG4 function and its therapeutic applications.

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Crystallization and preliminary X-ray diffraction study of a chimaeric Fab′ fragment of antibody binding tumour cells

Cited by 23 publications

References 6 publications

Monolayers assembled from a glycolipid biosurfactant from Pseudozyma (Candida) antarctica serve as a high-affinity ligand system for immunoglobulin G and M

Monolayers assembled from a glycolipid biosurfactant from Pseudozyma (Candida) antarctica serve as a high-affinity ligand system for immunoglobulin G and M

The application of force microscopy to immunodiagnostic systems: imaging and biomolecular adhesion measurements

The Fab Conformations in the Solution Structure of Human Immunoglobulin G4 (IgG4) Restrict Access to Its Fc Region

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