Quantitative determination of surface concentration of protein with surface plasmon resonance using radiolabeled proteins

Stenberg, Esa; Persson, Björn; Roos, Håkan; Urbaniczky, Csaba

doi:10.1016/0021-9797(91)90284-f

Cited by 1,012 publications

(730 citation statements)

References 21 publications

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“…[16] In our SPR set up, increased intensity correlates to DNA or OGNP adhesion, and loss of intensity is due to desorption of the chemical species from the surface. After addition of the DNA origami an increase of 3.29% of the intensity was observed, indicating adsorption of the nanostructure on the gold surface.…”

Section: Revised Manuscriptmentioning

confidence: 82%

DNA‐Origami‐Driven Lithography for Patterning on Gold Surfaces with Sub‐10 nm Resolution

et al. 2017

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The programmability [1] and self-assembly properties of DNA provides means of precise organization of matter at the nanoscale. [2] DNA origami allows the folding of DNA into twodimensional [3] and three-dimensional [4] structures, and has been used to organize biomolecules, [2b, e, 5] nanophotonic [2a, c, f, 6] and electronic [7] components with a resolution of 6 nm / pixel. [8] Two-dimensional DNA origami has been also used as a platform to organize other chemical [9] species that can then be placed on technologically relevant substrates. [2c, 10] Nevertheless, these approaches have only used the DNA nanostructure to hold the chemical species on the surface and, to the best of our knowledge, have never been utilized to immobilize nucleic acids patterns on surfaces with sub-10 nm resolution providing an enable 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 Submitted to 2 platform for potential applications such as multiplexed biochemical assays. [11] to the creation of metasurfaces [12] with potentially reconfigurable features. Revised ManuscriptHerein we report on the use of a two-dimensional DNA origami as a template to covalently attach DNA with a pre-programmed pattern on a surface (see Scheme 1). The method utilizes the incorporation of modified staple strands in programmed positions of the DNA origami (DNA origami stamp), acting as DNA ink. Once the DNA origami is immobilized on the surface, the modified staples can react with the surface creating a defined DNA pattern (Stamping step). The pattern can then be exposed upon denaturation of the DNA origami stamp (Unmasking step), allowing the non-bound staples to be rinsed off of the surface. As a proof-of-principle of this methodology, we have created a linear pattern of thiolmodified DNA ink on gold surfaces. The formation of the linear pattern was revealed by the successful formation of bead-on-a-string-like structures (here named "chains" for simplicity) composed of gold nanoparticles conjugated with thiol-oligonucleotides (OGNP) that are hybridized to the DNA ink pattern (Development step). The linear pattern provided a direct evidence of the stamping process and was chosen as a simple geometry that can be statistically analysed in our experimental setup. Montecarlo Simulations have been used for better understanding of our statistical results and to determine key elements governing the process that can be used for future optimization of pattern information transfer with DNA origami stamp methodology. Furthermore, we have studied the development of more complex patterns using Montecarlo Simulations.We demonstrate that our approach can be employed to form DNA patterns with sub-10 nm resolution to flat gold surfaces, an unsolved goal to date. This methodology can thus be extended to other surfaces utilizing different covalent strategies. [10b, 13] M...

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Section: Revised Manuscriptmentioning

confidence: 82%

DNA‐Origami‐Driven Lithography for Patterning on Gold Surfaces with Sub‐10 nm Resolution

et al. 2017

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“…The construct differs in that the L chain Cys at residue 57 (L55 in Kabat numbering) is at the end of, rather than before, the CDR L2, and the H chain Cys at position 106 (108 in Kabat numbering) is positioned four residues earlier in the sequence. The H-chain Cys is at a position where an Asp required for a salt-bridge to Arg-H98 (94 in Kabat numbering) is often found, so this was compensated for by altering the next residue to Asp, a mutation previously shown to retain full antigen-binding activity in Fab [15]. The position of the disulphide bond is closer to the binding-site than in the Jung et al design [3] but not as close as in the design of Glockshuber et al [2], which involves CDR H3 residues.…”

Section: Discussionmentioning

confidence: 99%

“…In many antibodies, the residue Asp-H106 is a partner in a highly-conserved saltbridge with Arg-H98. However, previous mutagenesis experiments [15] had shown that replacement of the next residue, Tyr-H107, with Asp gave an Fab with full antigen-binding activity. In order to retain an acidic group in this location as a potential salt-bridge partner for Arg-H98, this replacement was made as well.…”

Section: Design and Production Of The Ds-scfvmentioning

confidence: 94%

“…The antigen used was a conjugate of the Salmonella O-chain and bovine serum albumin and it was attached to the dextran-gold surface, using the amine coupling kit supplied by the manufacturer, in 10 mM Na acetate, pH 4.5, at concentrations and contact times that yielded about 200 RU of immobilized material. 1 RU corresponds to an immobilized protein concentration of 1 pg/mm 2 [15]. All measurements were performed at ambient temperature in 10 mM HEPES, pH 7.4, 100 mM NaC1, 3.3 mM EDTA at a flow rate of 5 pl/min and protein concentrations between 1.8 and 4.6 pM.…”

Section: Immunological Analysesmentioning

confidence: 99%

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Thermal stabilization of a single‐chain Fv antibody fragment by introduction of a disulphide bond

et al. 1995

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A disulphide bond was introduced into a single-chain Fv form of the anticarbohydrate antibody, Se155-4 by replacing Ala-L57 of the light chain and Asp-H106 of the heavy chain with cysteines, by site-directed mutagenesis. To maintain the saltbridge from the latter residue to Arg-H98, Tyr-107 was also altered to Asp. The resulting ds-scFv was shown to retain full antigen-binding activity, by enzyme immunoassay and surface plasmon resonance analysis of binding kinetics. Compared with the parent scFv, the disulphide bonded form was shown to have enhanced thermal stability, by Fourier transform IR spectroscopy. The Tm was raised from 60°C to 69°C. The ds-scFv form thus combines the stable monomeric form of the disulphide form wifh the expression advantages of the scFv.

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“…The main method was the QCM-D technique (13-15) but complementary SPR measurements (16)(17)(18) were also performed for two proteins. We measured the adsorption tendency of a number of proteins on a supported phospholipid bilayer, composed of electrically neutral eggPC lipids in the liquid crystalline state, supported on a SiO 2 surface.…”

Section: Introductionmentioning

confidence: 99%

Protein Adsorption on Supported Phospholipid Bilayers

Glasmästar

Larsson

Höök

et al. 2002

Journal of Colloid and Interface Science

294

283

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Quartz crystal microbalance with dissipation (QCM-D) measurements were used to investigate the adsorption of human fibrinogen, human serum albumin, bovine hemoglobin, horse heart cytochrome c, human immunoglobulin (hIgG), and 10% fetal bovine serum on supported bilayers of egg-phosphatidylcholine (eggPC) lipids. For comparison the adsorption of fibrinogen and hIgG to eggPC bilayers was also studied with surface plasmon resonance (SPR). The supported bilayers were formed in situ by vesicle adhesion and spontaneous fusion onto a SiO 2 surface. The supported lipid bilayer is highly protein resistant: The irreversible adsorption measured with the QCM-D technique was below the detection level, while reversible protein adsorption was detected for all the proteins in the range 0.3-4% of the saturation coverage on a hydrophobic thiol monolayer on gold. The adsorbed amounts were slightly higher for the SPR measurements. Possible mechanisms for the protein resistance of eggPC bilayers are briefly discussed. C 2002 Elsevier Science

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Quantitative determination of surface concentration of protein with surface plasmon resonance using radiolabeled proteins

Cited by 1,012 publications

References 21 publications

DNA‐Origami‐Driven Lithography for Patterning on Gold Surfaces with Sub‐10 nm Resolution

DNA‐Origami‐Driven Lithography for Patterning on Gold Surfaces with Sub‐10 nm Resolution

Thermal stabilization of a single‐chain Fv antibody fragment by introduction of a disulphide bond

Protein Adsorption on Supported Phospholipid Bilayers

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