Screening of Aptamers and Their Potential Application in Targeted Diagnosis and Therapy of Liver Cancer Introduction to Aptamers Aptamers are short, single-stranded DNA or RNA oligonucleotides (typically 20-100 nucleotides) that bind to specific target molecules with high affinity and specificity, similar to antibodies. They are often called "chemical antibodies" but offer advantages including: Ease of synthesis and modification Low immunogenicity Enhanced tissue penetration Thermal stability Lower production costs Screening Methods for Liver Cancer-Specific Aptamers 1. SELEX Technology Systematic Evolution of Ligands by Exponential Enrichment (SELEX) is the primary method for aptamer selection: Key adaptations for liver cancer: Cell-SELEX: Using live hepatoma cells (e.g., HepG2, Huh7) as targets Tissue-SELEX: Employing liver cancer tissue specimens In vivo SELEX: Direct screening within animal models Automated SELEX: High-throughput screening platforms 2. Target-Specific SELEX Variations Protein-based SELEX: Against liver cancer biomarkers (AFP, GPC3, etc.) Whole-cell SELEX: For cell surface epitope targeting Toggle SELEX: For cross-reactivity across different liver cancer cell types Applications in Liver Cancer Diagnosis 1. Imaging and Detection Molecular Imaging: Radiolabeled aptamers for PET/CT imaging Fluorescent Aptamers: For intraoperative guidance and tumor margin identification MRI Contrast Agents: Aptamer-conjugated nanoparticles for enhanced imaging 2. Biosensor Development Electrochemical Sensors: For detecting circulating tumor cells Colorimetric Assays: Point-of-care testing for…
Of course. Aptamers, often called "chemical antibodies," are single-stranded DNA or RNA oligonucleotides that bind to specific target molecules (proteins, small molecules, cells, etc.) with high affinity and specificity. Their unique properties—ease of synthesis, stability, low immunogenicity, and small size—make them powerful tools across numerous fields. Here is a comprehensive overview of the key applications of aptamers, categorized by field: 1. Therapeutics & Medicine Aptamers are developed as targeted drugs, often competing with monoclonal antibodies. Targeted Cancer Therapy: Aptamers can deliver toxic drugs, siRNAs, or radioisotopes directly to cancer cells by binding to overexpressed surface markers (e.g., nucleolin, PSMA). This minimizes damage to healthy tissues. Antidotes and Antagonists: The only FDA-approved aptamer to date, Pegaptanib (Macugen), is used for age-related macular degeneration (AMD). It binds and inhibits VEGF-165, a protein promoting abnormal blood vessel growth. Antiviral and Antimicrobial Agents: Aptamers can block viral entry (e.g., against HIV gp120, SARS-CoV-2 spike protein) or inhibit essential bacterial proteins, offering potential alternatives to traditional antibiotics. Anti-inflammatory and Autoimmune Diseases: Aptamers can neutralize inflammatory cytokines (e.g., TNF-α, IL-6) implicated in diseases like rheumatoid arthritis or psoriasis. Targeted Drug Delivery Systems: Aptamers conjugated to nanoparticles or liposomes create "smart" delivery vehicles that release their cargo only at the disease site. 2.…
The Core Principle of SELEX Think of SELEX as artificial selection or directed evolution for molecules. You start with a massive, diverse library of random nucleic acid sequences (typically 10^14 to 10^15 different molecules) and iteratively select for the tiny fraction that binds tightly to your target molecule. The Standard SELEX Process (Step-by-Step) The process is cyclical, typically requiring 5-15 rounds to enrich high-affinity binders. 1. Library Design & Synthesis: A synthetic library is created where each molecule has constant primer regions (for PCR amplification) flanking a central randomized region (20-60 nucleotides long). This creates a "pool of possibility" often called a combinatorial library. 2. Incubation (Binding): The library is incubated with the target molecule (e.g., a protein, small molecule, cell, or even a whole pathogen). Conditions (buffer, temperature, ionic strength) are controlled to favor specific binding. 3. Partitioning (Separation): This is the most critical step. The bound sequences (potential aptamers) must be efficiently separated from the unbound ones. Common methods: immobilizing the target on a column/beads, filtration through nitrocellulose filters (which trap protein-bound RNA/DNA), or capillary electrophoresis. 4. Elution (Recovery): The bound sequences are recovered, often by denaturing the complex (e.g., heating, changing pH, or using denaturants). 5. Amplification: The recovered sequences are amplified using PCR (for DNA aptamers) or Reverse Transcription-PCR (for…
1. Synthetic & Chemical Origin Fully In Vitro Production: Unlike biological antibodies produced in living cells (mice, rabbits, CHO cells), Adaptamers are synthesized chemically in a controlled laboratory setting. Precise Chemical Structure: Their composition, structure, and modifications are defined at the atomic level, leading to perfect batch-to-batch consistency. No Biological System Required: Production is independent of cell culture, fermentation, or animal use, making it more scalable and ethically straightforward. 2. Adaptive & Engineered Binding "Adapta-" Implies Engineering: The name suggests the molecule is engineered or evolved in vitro (like aptamers via SELEX) to bind with high affinity and specificity to a chosen target—proteins, small molecules, cells, etc. Tailor-Made: Their binding sites are not constrained by biological immune system mechanisms. They can be designed against targets that are poorly immunogenic, toxic, or highly conserved. 3. Core Molecular Scaffold (Likely Nucleotide or Peptide-Based) While unspecified, the most probable backbones are: Nucleic Acid-Based (like advanced aptamers): Composed of DNA or RNA, but heavily chemically modified (e.g., with 2'-fluoro, 2'-O-methyl, locked nucleic acids (LNA), or entirely synthetic bases) to confer nuclease resistance and enhanced stability. Peptide-Based (like peptidomimetics or constrained peptides): Composed of non-natural amino acids or cyclic structures for rigidity and protease resistance. Hybrid or Other Polymer: Could be a fusion of chemistries (e.g., oligonucleotide with a peptide-like side chain) for…
What are Aptamers? Aptamers are single-stranded DNA or RNA molecules obtained through in vitro selection techniques (such as SELEX) from synthetic, randomized nucleic acid libraries. They can bind to target molecules (such as proteins, small molecules, cells, etc.) with high affinity and specificity through unique three-dimensional structures, earning them the titles "chemical antibodies" or "nucleic acid antibodies." Key Characteristics Chemical Nature Mostly single-stranded DNA or RNA, producible via chemical synthesis, offering good stability and ease of modification or labeling. Binding Mechanism Form hydrogen bonds, electrostatic interactions, etc., through spatial conformations (e.g., hairpins, pockets, bulges) to recognize targets with atomic-level precision. High Affinity & Specificity Capable of distinguishing structurally similar molecules (e.g., caffeine vs. theophylline), with dissociation constants (Kd) reaching nanomolar (nM) or even picomolar (pM) levels. Strong Stability Resistant to high temperatures, reversible denaturation, and less susceptible to damage in acidic/basic environments (especially DNA aptamers), making them suitable for complex applications. Advantages Compared to Antibodies Feature Aptamers Antibodies (Traditional Protein Antibodies) Production Time In vitro selection (weeks), chemical synthesis, high batch consistency Animal immunization or cell culture (months), significant batch variability Stability Heat-resistant, easy storage, transportable at room temperature Prone to denaturation, requires cold storage Modification Flexibility Easy labeling with fluorescent dyes, isotopes, etc. Modifications may affect structure Immunogenicity…
Aptamers are nucleic acid sequences that specifically bind with target molecules and are vital to applications such as biosensing, drug development, disease diagnostics, etc. The traditional selection procedure of aptamers is based on the Systematic Evolution of Ligands by an Exponential Enrichment (SELEX) process, which relies on repeating cycles of screening and amplification. With the rapid development of aptamer applications, RNA and XNA aptamers draw more attention than before. But their selection is troublesome due to the necessary reverse transcription and transcription process (RNA) or low efficiency and accuracy of enzymes for amplification (XNA). In light of this, we review the recent advances in aptamer selection methods and give an outlook on future development in a non-SELEX approach, which simplifies the procedure and reduces the experimental costs. We first provide an overview of the traditional SELEX methods mostly designed for screening DNA aptamers to introduce the common tools and methods. Then a section on the current screening methods for RNA and XNA is prepared to demonstrate the efforts put into screening these aptamers and the current difficulties. We further predict that the future trend of aptamer selection lies in non-SELEX methods that do not require nucleic acid amplification. We divide…
KMD Bioscience offers comprehensive aptamer discovery services that cover the full workflow—from nucleic acid aptamer design and synthesis, to multi-target SELEX screening, to final affinity and specificity validation. With advanced laboratory platforms and experienced scientists, we provide reliable and efficient one-stop aptamer solutions for diagnostics, therapeutics, biosensors and research applications. Nucleic Acid Aptamer Introduction Aptamers are single-stranded nucleic acid molecules (DNA or RNA) that fold into stable three-dimensional structures capable of binding specific targets with high affinity. Compared with antibodies, aptamers offer several advantages, including small molecular size, low immunogenicity, chemical synthesis, batch consistency and flexible modification. They can bind diverse targets such as proteins, peptides, cells, small molecules and ions, making them valuable tools in biotechnology and analytical applications. DNA Aptamer: DNA aptamers are aptamers composed of deoxyribonucleic acid (DNA). They usually have a double helix structure and are composed of four bases (adenine A, thymine T, guanine G, cytosine C) connected by phosphodiester bonds. They are single-stranded oligonucleotide sequences, typically with 56-120 bases, that bind the target sequence efficiently by recognizing specific spatial structures. DNA aptamers are widely used in biological analysis, biomedicine, and other fields because of their stability and easy chemical modification. RNA Aptamers: RNA aptamers are aptamers…
1. What is metal ion aptamer screening? A: The aptamer is a small nucleic acid folded in a three-dimensional conformation to make the aptamer a specific binding target. The target has a variety of species, either proteins small molecules, or metal ions. In contrast to antibodies, aptamers enable in vitro selection with low immunogenicity. Among them, heavy metal, as the most toxic metal pollutant, can pollute the natural environment, and detecting heavy metal pollution has become an important task. At the same time, heavy metals are also classified as trace elements because of their low content. Therefore, developing an accurate and sensitive heavy metal detection method is very important to ensure human and environmental safety. Aptamers as biological probes show high binding affinity that can be directly converted to high detection sensitivity. On the other hand, high selectivity and stability enable it to detect various targets, especially metal ions. Since identifying aptamers for metal ions, aptamer-based sensors and detection methods have become possible and provide new methods for detecting metal ions, including electrochemical, colorimetric, and fluorometry. KMD Bioscience has been studying in the field of nucleic aptamers for a long time, which can provide customers with metal ion aptamers with excellent performance to help customers better…
Due to its high specificity, strong affinity, excellent stability, and ease of preparation and labeling, the screening techniques for aptamers in fields such as cells, fungi, proteins, and small molecules have become increasingly sophisticated. Due to the small molecular weight and single binding site of metal ions, screening methods need to be specially designed. Current research mainly focuses on heavy metal ion aptamers, such as Pb2+, Cd2+, and Hg+. KMD Bioscience provides metal ion nucleic acid aptamer screening services through affinity chromatography SELEX and graphene oxide SELEX, based on the selenium library screening technology. When conducting metal ion adaptation screening, there are certain requirements for the sample, and the metal ion sample should have high purity to reduce the interference of impurities in the screening process. The concentration of metal ions should be moderate, neither too high nor too low, to ensure effective binding during the screening process. Metal ion samples should be kept stable to avoid chemical changes or degradation during the screening process. Relatively speaking, selex library should have sufficient sequence diversity to cover possible adapter sequences. The length of oligonucleotide sequences should be moderate to form stable structures and effectively bind with metal ions. The metal ion sample and…
Metal Ion Nucleic Acid Aptamer Screening Service Workflow Step Service Content Timeline Step 1: Screening of nucleic acid aptamers (1) Customer provides screening targets. (2) The adapter library is fixed on an affinity chromatography column and incubated with metal ions injected into the column. (3) Adaptation library screening and enrichment: PCR amplification enrichment+transcription+gel running recovery, usually 6-10 rounds. (4) Screening products for NGS sequencing. (5) Delivery: 5-15 adapter sequences, experimental report, raw data (including NGS sequencing raw data and gel electrophoresis) 10-15 weeks Step2：Synthesis of aptamers and determination of affinity (optional) (1) Synthesize aptamers based on sequences. (2) Affinity determination of adapter and target protein, KD determination by BLI or SPR. (3) Delivery: Experimental report, raw data
