Method, surface, particle and kit for the detection of analytes in samples
AssigneesTechnische UniversitäT Dresden, UniversitäT Leipzig
InventorsGerhard RÖDEL, Kai OSTERMANN, Julia DÖRING, Christian DAHMANN, Tilo POMPE, David RETTKE, Steve MARTIN
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Abstract
The invention relates to a method, to a surface, to a particle, and to a kit for the detection of low molecular weight analytes such as crop protection agents in samples. In particular, the invention relates to a method for the detection of glyphosate through protein-functionalised surfaces and functionalised particles by means of reflection interference contrast microscopy (RICM).
Claims20
A method for the detection of analytes comprising the steps of:
- providing a surface with an immobilised analyte binding partner,
- contacting the analyte binding partner with a sample containing the analyte, wherein the analyte interacts with the analyte binding partner,
- contacting the analyte binding partner with a competitor, wherein the competitor is immobilised on a particle and interacts with the analyte binding partner,
- detecting the competitors bound to the analyte binding partner,
- wherein the analyte is glyphosate,
- wherein the analyte binding partner has the active centre of the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) as the analyte binding site, and
- wherein the particle is designed to be deformable.
A surface having an analyte binding partner, wherein the analyte binding partner is a fusion protein having a hydrophobin domain and a 5-enolpyruvylshikimate-3-phosphate synthase domain, wherein the surface is designed to be transparent at least in the wavelength range from 400 nm to 600 nm.
A particle having an immobilised competitor, wherein the competitor is immobilised on the particle via a linker, wherein the particle is designed to be deformable, wherein the competitor is selected from phosphoenolpyruvate, phosametine, substrate analogues of the enzyme 5-enolpyruvylshikimate-3-phosphate synthase or glyphosate,
- wherein the linker has a contour length of 5-200 Å and/or a degree of polymerisation of 1-70.
A kit, comprising:
- at least one immobilised analyte binding partner or one analyte binding partner and a hydrophobin, wherein the analyte binding partner is designed to interact with an analyte and is immobilised on a surface,
- wherein the analyte binding partner is a fusion protein having a hydrophobin domain and a 5-enolpyruvylshikimate-3-phosphate synthase domain, and
- wherein the surface is designed to be transparent at least in the wavelength range from 400 nm to 600 nm;
- and
- at least one particle, having an immobilised competitor,
- wherein the competitor is immobilised on the particle via a linker,
- wherein the particle is designed to be deformable.
Dependencies are inferred from the claim text.