What is Capture-SELEX? Unlike traditional SELEX where the target is immobilized, Capture-SELEX immobilizes the initial DNA library itself via a short complementary "capture" sequence. The key target molecule is free in solution. Binding occurs when an aptamer candidate in the library binds to the target, causing a structural change that releases it from the immobilization surface. This approach offers distinct advantages: Ideal for small molecules and proteins: Especially targets that are difficult to immobilize without affecting their structure. Minimizes non-specific binding: Selection pressure is purely for target-induced structure formation/release. Enriches for structure-switching aptamers: Resulting aptamers often undergo conformational change upon binding, making them excellent for biosensor development. Typical Capture-SELEX Screening Service Workflow A professional service provider will manage this complex, iterative process from start to finish. Here’s what you can expect: Phase 1: Project Design & Library Preparation Consultation & Target Specification: Defining target properties, desired affinity (Kd), specificity (against which counter-targets), and buffer conditions. Customized Library Design: Designing a single-stranded DNA library (10^14 - 10^15 unique sequences) with: A central random region (e.g., 30-50 nucleotides). Fixed primer regions for PCR amplification. A capture sequence region complementary to an immobilized oligonucleotide. Immobilization Matrix Preparation: Coupling the complementary "capture" oligonucleotides to a solid support (e.g., magnetic beads, chromatography resin). Phase 2: The Iterative Selection (SELEX) Cycles…
What is Filter Membrane Binding SELEX? SELEX (Systematic Evolution of Ligands by EXponential enrichment) is the standard method for discovering high-affinity, specific nucleic acid aptamers. The Filter Membrane Binding variant is one of the most classic and robust SELEX techniques. Core Principle: It leverages a nitrocellulose or mixed cellulose ester filter membrane, which irreversibly binds proteins and other macromolecules but allows short, unbound single-stranded DNA or RNA oligonucleotides to pass through. The Selection Mechanism: During each selection round, the target molecule (e.g., a protein) is immobilized on the filter. An immense library of random oligonucleotides (10^13 - 10^15 unique sequences) is applied. Only sequences that bind to the target are retained on the filter with it. Unbound sequences are washed away. The bound aptamer candidates are then eluted, amplified by PCR (or RT-PCR for RNA), and used as the enriched library for the next round. Key Features of the Service A professional service will typically offer: Target Flexibility: Optimal for purified proteins (recombinant or native), protein complexes, viruses, and even some small molecules if conjugated to a carrier protein. Counter-SELEX: A critical step to ensure specificity. The enriched library is passed through a filter bound to non-target molecules (e.g., related proteins, cell lysates, immobilization matrix) to subtract cross-reactive binders. High-Throughput…
What is SELEX and What are Aptamers? Aptamers: Often called "chemical antibodies," they are short, single-stranded DNA or RNA oligonucleotides that fold into specific 3D shapes to bind with high affinity and specificity to a target molecule (e.g., a viral protein, whole bacterium, or parasite surface marker). SELEX (Systematic Evolution of Ligands by EXponential enrichment): This is the iterative combinatorial chemistry process used to discover aptamers from a vast random library (10^14-10^15 unique sequences). It involves repeated cycles of: 1) Binding the library to the target, 2) Separating bound from unbound sequences, 3) Amplifying the bound sequences, and 4) Starting a new, enriched cycle. Core Components of a Pathogen SELEX Service A professional service will typically manage the entire pipeline: 1. Project Design & Target Preparation: Consultation: Defining the precise target (e.g., whole inactivated SARS-CoV-2, Salmonella outer membrane protein, Plasmodium lysate). Counter-SELEX: A critical step for pathogen specificity. The process is run against related non-targets (e.g., host cells, non-pathogenic bacterial strains) to filter out cross-reactive aptamers, ensuring the final aptamers distinguish between pathogen and non-pathogen. 2. The SELEX Execution: Performing multiple (usually 8-15) rounds of the selection process under optimized conditions (buffer, temperature, washing stringency). 3. Next-Generation Sequencing (NGS) & Bioinformatics: After the final rounds, the enriched pool is sequenced using NGS. Bioinformatic analysis identifies sequence…
Tissue-SELEX Aptamer Screening Service This refers to a specialized contract research service that uses Systematic Evolution of Ligands by EXponential Enrichment (SELEX) to discover aptamers that specifically bind to molecular targets within a complex tissue environment, rather than purified proteins or isolated cells. Core Concept & Key Differentiator While Cell-SELEX uses whole live cells as targets, Tissue-SELEX advances the complexity by using: Tissue sections (fresh, frozen, or FFPE - Formalin-Fixed Paraffin-Embedded) Tissue homogenates Tissue-specific extracellular matrix (ECM) components The goal is to find aptamers that recognize targets in their native, histological context, preserving post-translational modifications and local microenvironments. This is crucial for developing reagents for histopathology and tissue-specific targeting. Typical Workflow Target Preparation: Provider prepares or client supplies well-characterized tissue sections (often on glass slides). Counter-selection tissues (e.g., healthy vs. diseased, organ A vs. organ B) are critical. SELEX on Tissue: The oligonucleotide library is incubated directly on the tissue section. After washing, bound sequences are eluted, often by laser capture microdissection of bound areas or direct extraction. Amplification & Iteration: Recovered sequences are amplified (PCR) and used for subsequent selection rounds, with increasing stringency. Sequencing & Analysis: High-throughput sequencing (NGS) identifies enriched sequence families. Validation: Top candidates are synthesized and validated via: Tissue Staining: Fluorescently-labeled aptamers used like antibodies in immunohistochemistry (IHC). Specificity: Testing on…
What is a Stem Cell Aptamer Screening Service? It is a contract research service where a specialized lab uses Systematic Evolution of Ligands by EXponential Enrichment (SELEX) to discover and develop DNA or RNA aptamers that bind with high affinity and specificity to a target of your choice related to stem cells. Aptamers are often called "chemical antibodies." They are short, single-stranded oligonucleotides that fold into unique 3D shapes, allowing them to bind to targets like proteins, small molecules, or even whole cells. Core Targets for Stem Cell Applications The service can be tailored to screen for aptamers against: Specific Cell Surface Markers: (e.g., CD34, CD133, SSEA-4, TRA-1-60) for identification and isolation. Whole Live Stem Cells: To get aptamers that recognize the unique molecular signature of a specific stem cell type (e.g., mesenchymal stem cells, cancer stem cells, pluripotent stem cells). Differentiation State-Specific Targets: To distinguish between pluripotent, progenitor, and fully differentiated cells. Specific Stem Cell-Derived Products: (e.g., exosomes, vesicles). Typical Workflow of the Service A professional service provider will guide you through these stages: Phase Description Your Input 1. Project Design Defining the target (specific protein, cell line, primary cells), counter-selection cells (to ensure specificity), and desired aptamer properties (e.g., Kd, nuclease resistance). Provide target cells, control cells, and…
What is a Bacterial Aptamer Screening Service? It is a specialized contract research service where a provider uses Systematic Evolution of Ligands by Exponential Enrichment (SELEX) to discover and develop single-stranded DNA or RNA aptamers that bind with high affinity and specificity to a bacterial target. The target can be: Whole bacterial cells (e.g., E. coli O157:H7, Salmonella typhimurium). Specific bacterial components (e.g., surface proteins like pili, flagella, capsular polysaccharides, secreted toxins). Key virulence factors (e.g., endotoxins like LPS). The resulting aptamers are powerful recognition elements for diagnostics, therapeutics, and research. Core Steps in the Service Pipeline A typical full-service offering includes: 1. Project Design & Target Preparation: Consultation: Defining the goal (e.g., detection of a specific strain, therapeutic neutralization). Target Choice: Deciding between whole cells (for broad detection) or purified components (for precise targeting). Counter-SELEX: Using related non-target cells (e.g., non-pathogenic strain) to eliminate cross-reactive aptamers and ensure specificity. 2. Library Synthesis & SELEX Process: Library Design: Using a random-sequence oligonucleotide library (typically ~10^14 different molecules). Selection Rounds (8-15 cycles): Iteratively incubating the library with the target, washing away unbound sequences, eluting the bound ones, and amplifying them via PCR (for DNA) or RT-PCR (for RNA). Monitoring: Using quantitative PCR or flow cytometry to track enrichment progress. 3. Next-Generation Sequencing (NGS) & Bioinformatics:…
What is the Core Service? The service provider uses an iterative, in vitro selection process called SELEX (Systematic Evolution of Ligands by EXponential Enrichment) to screen vast random oligonucleotide libraries (10^14 - 10^15 unique sequences) against your target protein. The output is a set of characterized aptamer sequences that bind to the viral capsid. Standardized Screening Workflow A professional service will manage this entire pipeline: 1. Project Design & Target Preparation: Target Discussion: Defining the specific capsid protein (e.g., HIV-1 CA, HBV core, SARS-CoV-2 N), its form (full-length, domain, assembled capsid/nucleocapsid), and purity. Target Immobilization: The protein is often immobilized on a solid support (beads, plate) to facilitate separation of bound/unbound sequences. Some services offer solution-phase or capillary electrophoresis (CE-SELEX) methods for higher stringency. 2. SELEX Selection Rounds (Cycles 5-15): Incubation: The oligonucleotide library is incubated with the target. Partitioning: Unbound sequences are washed away; bound sequences are retained. Elution: Bound aptamers are eluted (e.g., by heating, denaturing agents). Amplification: Eluted aptamers are amplified by PCR (for DNA) or RT-PCR (for RNA). Purification: The amplified pool is purified for the next selection round. Counter-Selection: To ensure specificity, the pool is often passed through a negative control (e.g., irrelevant protein, cell lysate) to remove non-specific binders. 3. Sequencing & Identification: High-Throughput…
What is the Service? It's the process of using SELEX (Systematic Evolution of Ligands by EXponential Enrichment) to identify single-stranded DNA or RNA aptamers that can bind to a target cytokine. The service takes you from target selection to delivering validated aptamer candidates. Standard Workflow (What the Provider Does) Project Scoping & Target Preparation: Target: You specify the cytokine (e.g., TNF-α, IL-6, IFN-γ). The provider may require you to supply the purified, recombinant protein or offer to procure/produce it. Counter-SELEX: A critical step to ensure specificity. The provider will use related proteins (e.g., other cytokines, serum proteins) to eliminate aptamers that bind non-specifically. Library Design & SELEX Cycle: Starts with a vast random oligonucleotide library (10^14 - 10^15 unique sequences). Iterative rounds (8-15+) of: Binding: Incubating the library with the target cytokine. Partitioning: Separating bound from unbound sequences (e.g., via immobilization on beads, filters, or capillary electrophoresis). Amplification: PCR (for DNA) or RT-PCR (for RNA) to enrich the binding sequences. Stringency Increase: Gradually increasing washing rigor and introducing counter-selection to drive selection of high-affinity, specific binders. Next-Generation Sequencing (NGS) & Bioinformatics: After the final rounds, the enriched pool is sequenced using NGS. Bioinformatics tools analyze the data to identify enriched sequence families, consensus motifs, and predict secondary structures.…
What is an Aptamer? An aptamer is a short, single-stranded oligonucleotide (DNA or RNA) that folds into a unique 3D structure, allowing it to bind to a specific target molecule (like a protein) with similar specificity to an antibody. They are often called "chemical antibodies." Why Use a Screening Service Instead of In-House Development? Expertise & Equipment: The screening process (SELEX) requires specialized skills, robotics, and next-generation sequencing (NGS) infrastructure. Time & Cost Efficiency: Outsourcing can be faster and more cost-effective than setting up a new, complex pipeline. Higher Success Rate: Experienced providers have optimized protocols for difficult targets (e.g., membrane proteins, toxic proteins). The Core Process: SELEX The standard method is SELEX (Systematic Evolution of Ligands by EXponential Enrichment). A professional service will offer advanced variants of this process. A Typical Service Workflow: Project Consultation & Design: Target Characterization: Discussion about your protein (purified? membrane-bound? post-translational modifications?). Selection Strategy: Choosing the best SELEX method (e.g., Capillary Electrophoresis-SELEX (CE-SELEX) for very high affinity, Cell-SELEX for cell-surface targets, Toggle-SELEX for cross-species specificity). Counter-Selection: Designing the process to avoid binding to non-target proteins (e.g., carrier proteins, related isoforms). Library Synthesis & Preparation: Creation of a vast random oligonucleotide library (typically 10¹³ - 10¹⁵ unique sequences). The Selection Rounds (Cycles of SELEX): Binding: Incubating the library with the…
Core Technology: SELEX The foundation of all these services is the SELEX process, an in vitro method to select aptamers from a vast random library (typically 10^13 - 10^15 unique sequences). The library is incubated with the target, unbound sequences are washed away, and bound sequences are eluted and amplified by PCR (for DNA) or RT-PCR (for RNA). This cycle is repeated 8-15 times to enrich for the tightest binders. Services for Protein Targets This is the most common application, as aptamers are often touted as "chemical antibodies." 1. Standard Protein SELEX: Target: Purified, recombinant proteins (e.g., cytokines, receptors, enzymes, viral capsids). Key Considerations: Protein Purity & Conformation: Critical for success. Services often require >90% purity and verification of native folding. Immobilization: The protein is usually immobilized on beads (e.g., streptavidin/biotin, Ni-NTA/His-tag) to facilitate partitioning. Some services offer solution-phase SELEX to avoid conformation changes. Counter-Selection: To ensure specificity, libraries are pre-incubated with related proteins or the immobilization matrix to subtract non-specific binders. 2. Specialized SELEX for Complex Proteins: Membrane Protein SELEX: For receptors and channels. Requires special handling (e.g., use of nanodiscs, detergent micelles, or whole cells overexpressing the target). Post-Translationally Modified Protein SELEX: For targets where phosphorylation, glycosylation, etc., are essential for function. 3. Cell-SELEX (for Cell-Surface…
