A Method of Identifying or Producing an Aptamer for a Denatured Peptide or Protein

Simple SummaryContent extracted from patent full text and abstract with AI.

This patent introduces a method for identifying or producing aptamers (short nucleic acid molecules that bind targets with high specificity) that specifically recognize denatured forms of peptides or proteins—such as those found in Western blot assays. The process involves a specialized SELEX (Systematic Evolution of Ligands by Exponential Enrichment) protocol where candidate aptamers are selected and amplified based on their ability to bind denatured proteins separated on membranes. The method also enables chemical modification of aptamers via click chemistry to enhance binding specificity or enable direct detection with labels.

Use CasesContent extracted from patent full text and abstract with AI.

• Improving sensitivity and specificity in Western blotting assays as aptamer-based alternatives to antibodies.
• Detection and quantification of specific denatured proteins or peptides in research, diagnostics, or proteomics.
• Identifying cancer biomarkers or disease-specific proteins in denatured form for diagnostic purposes.
• Development of custom aptamer reagents for protein analysis in academic or industrial laboratories.
• Direct labeling of aptamers for streamlined one-step protein detection, eliminating secondary antibody steps in assays.

BenefitsContent extracted from patent full text and abstract with AI.

• Enables highly specific and reproducible detection of denatured proteins or peptides, outperforming conventional antibodies in Western blots.
• Chemically synthesized aptamers are cheaper, easier to modify, and less variable between batches compared to antibodies.
• Flexible chemical modification (via click chemistry) allows integration of fluorescent or other reporter tags for direct detection.
• Reduces assay complexity by potentially eliminating the need for secondary reagents or multiple detection steps.
• Facilitates selection of aptamers directly under the conditions of the intended analytical application (e.g., on Western blots), ensuring optimal performance.
• Applicable to a wide range of protein targets, including single proteins or complex mixtures such as whole cell lysates.

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Patent Abstract

The invention relates to a method of identifying or producing an aptamer, the method comprising the steps of: a) Providing a candidate mixture of nucleic acids; b) Contacting the candidate mixture with a target sample; c) Partitioning nucleic acids that bind to the target sample from those that do not bind; d) Amplifying the binding nucleic acids to produce a population of nucleic acids that bind the target sample; e) Single strand displacement of the amplified nucleic acids, f) optionally repeating the steps a) to e), thereby identifying or producing an aptamer, wherein the target sample provides one or more peptides and/or proteins in a denatured form.