An aptamer library is a diverse pool of nucleic acid sequences (DNA or RNA) from which specific aptamers—short oligonucleotides that bind to target molecules with high affinity—can be selected. Constructing a high-quality library is the foundation of aptamer screening technologies like SELEX. 2. Key Components of an Aptamer Library Randomized Region The central portion of the aptamer, typically 20–60 nucleotides, is randomized to generate diversity. Example: N20–N40 where N = A, T/U, G, or C. The diversity determines the probability of finding high-affinity binders. Flanking Constant Regions Short sequences (~15–25 nt) at both ends of the randomized region. Functions: Primer binding sites for PCR amplification. Stability and structural constraints. Overall Length Usually 40–100 nucleotides, balancing structural complexity and amplification efficiency. 3. Steps of Library Construction Design of Oligonucleotides Include random regions flanked by known primer sequences. Example structure:5'-[Primer]-N40-[Primer]-3' Chemical Synthesis Use solid-phase DNA/RNA synthesis to generate the oligonucleotides. Random nucleotides are incorporated using a controlled mixture of A, T/U, G, C. Amplification (for DNA libraries) PCR amplifies the synthesized sequences. RNA libraries require in vitro transcription from DNA templates. Purification Remove truncated or incomplete sequences. Methods: PAGE purification or HPLC. Quality Control Ensure correct length, diversity, and absence of biases.…
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…
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 an Antibody Aptamer Screening Service? It is a specialized contract research service where a biotechnology company uses SELEX (Systematic Evolution of Ligands by EXponential Enrichment) or advanced variations of it to discover and develop aptamers that bind with high affinity and specificity to a target antibody. Antibody: A large, Y-shaped protein produced by the immune system to identify and neutralize pathogens. Aptamer: A short, single-stranded DNA or RNA oligonucleotide (or a modified derivative) that folds into a specific 3D structure, enabling it to bind to a target molecule with antibody-like specificity. Often called "chemical antibodies." The goal of the service is to provide clients with synthetic, recombinant-like binding molecules as alternatives or complements to traditional monoclonal antibodies. Why Screen for Aptamers Against Antibodies? Aptamers offer distinct advantages, making them attractive for various applications: Anti-Drug Antibody (ADA) Detection: Develop aptamer-based assays to detect and quantify ADAs in clinical trials for biotherapeutics. Diagnostic Tools: Create aptamer sensors (aptasensors) to detect specific antibody biomarkers for diseases (e.g., autoantibodies in autoimmune disorders). Therapeutic Neutralization: Discover aptamers that can bind and neutralize pathological antibodies (e.g., in autoimmune diseases like lupus or myasthenia gravis). Purification & Pull-Down: Use aptamers as ligands in chromatography or in assays to capture and isolate specific antibodies from complex…
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 Concept: What is SELEX? SELEX (Systematic Evolution of Ligands by EXponential Enrichment) is an iterative, in vitro selection process. It starts with a vast, random library of oligonucleotides (10^14 - 10^15 unique sequences) and, over multiple rounds, enriches for those that bind to the target. Standard Multi-Round SELEX Screening Service Workflow A full-service provider will typically manage the entire process, which can be broken down into key phases: Phase 1: Project Design & Target Preparation Target Consultation: Defining the target (e.g., protein, small molecule, cell, virus). Critical discussion of target purity, immobilization strategy, and selection conditions (buffer, temperature, counter-selection). Library Design: Selection of a random library (e.g., 40-nt random core with fixed primer sites). Options include DNA, RNA (requiring reverse transcription), or modified libraries (e.g., with 2'-F pyrimidines for nuclease resistance). Immobilization Strategy: The service provider will choose the best method: Immobilized Target: (Most common for proteins) Binding target to beads (streptavidin, Ni-NTA for His-tag) or columns. Counter-Selection: Using negative control surfaces (e.g., blank beads, related but undesired proteins) to subtract non-specific binders. Phase 2: The SELEX Cycle (Repeated 8-15 Rounds) This is the core iterative screening process. Each round consists of: Incubation: The oligonucleotide library is incubated with the target under defined conditions. Partitioning: Separation of…
What is an Aptamer? First, a quick reminder: Aptamers are short, single-stranded DNA or RNA oligonucleotides that bind to a specific target with high affinity and specificity. They are often called "chemical antibodies." The Core Service: SELEX (The Screening Process) The service revolves around executing a SELEX (Systematic Evolution of Ligands by EXponential enrichment) campaign. This is an iterative, in-vitro combinatorial chemistry process that screens a vast random library (10^14 - 10^15 unique sequences) to find the few that bind your target. A standard SELEX workflow includes: Library Design & Synthesis: Creating the initial random oligonucleotide pool. Incubation: The library is exposed to the target. Partitioning: Bound sequences are separated from unbound ones (the most critical step, varying by target type). Amplification: The bound sequences are amplified (usually by PCR for DNA, RT-PCR for RNA). Counter-Selection (Negative Selection): To increase specificity, the pool is exposed to non-target surfaces (e.g., immobilization matrix, related proteins) to remove non-specific binders. Repetition: Steps 2-5 are repeated for 8-15 rounds until a high-affinity pool is enriched. Cloning & Sequencing: The final pool is cloned, and individual aptamer sequences are identified via Next-Generation Sequencing (NGS). Bioinformatics & Analysis: NGS data is analyzed to identify candidate sequences, often clustered into families based on sequence/structure motifs. Characterization: Top candidates…
What is Classical SELEX? SELEX is an iterative, in vitro selection process used to isolate single-stranded DNA or RNA molecules (aptamers) that bind with high affinity and specificity to a target (e.g., a protein, small molecule, cell, or virus). The "classical" method refers to the original, well-established protocol involving: Incubation: A vast, random-sequence nucleic acid library (10^14 - 10^15 different sequences) is exposed to the target. Partitioning: Unbound sequences are washed away; bound sequences are retained. Elution: The bound sequences are recovered. Amplification: The recovered sequences are amplified by PCR (for DNA) or RT-PCR (for RNA). Repetition: This cycle (typically 8-15 rounds) is repeated, enriching the pool for the strongest binders. Components of a Classical SELEX Service A full-service provider typically manages the entire pipeline: 1. Project Design & Consultation Target Characterization: Discussing the target's properties (purity, stability, availability). Selection Strategy: Deciding on immobilization method (e.g., target immobilized on beads, or "counter-SELEX" to eliminate binders to the immobilization matrix or similar non-target molecules). Library Design: Choosing DNA or RNA, length of the random region (typically 20-60 nt), and fixed primer regions. 2. The SELEX Process Execution Library Synthesis: Chemical synthesis of the initial random library. Cycle Management: Performing the repetitive rounds of binding, washing, elution, and amplification under optimized buffer and stringency…
