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Nucleic Acid Aptamer Modification Service

TEK Biotech, with its profound experience in antibody research, has successfully delivered many antibody development achievements, constructed a comprehensive aptamer design framework, and accumulated a large amount of project practical experience in the process. TEK Biotech is dedicated to providing customers with high-quality nucleic acid adapter screening services and strongly supporting them in downstream research activities such as aptamer binding assays, modifications, aptamer conjugates, functional validation, and drug development targeting specific molecules. In addition, TEK Biotech also has the ability to provide one-stop services (from gene analysis synthesis to in vitro screening, aptamer binding assay, affinity determination, and the entire process of nucleotide adapter modification, including RNA and DNA nucleic acid adapter modification) to meet the diverse project needs of various types of customers. Through its professional aptamer design services, TEK Biotech provides aptamers that can quickly and accurately detect target molecules, resulting in a significant improvement in efficiency. Multiple hours of modifying groups are available for selection, and TEK Biotech ensures that every customer can obtain efficient and accurate nucleic acid adapter results, adding support to their research journey.

TEK Biotech provides SELEX screening technology services to customers, covering various types of samples (including but not limited to proteins, peptides, amino acids, and small molecule substances). The company's library has a capacity of up to 10 ^ 14 levels, which is sufficient to screen for high affinity nucleic acid aptamers targeting specific customer targets. By using various screening strategies such as magnetic bead SELEX, cell SELEX, and capture SELEX, highly specific nucleic acid aptamers can be identified after 6 to 10 rounds of rigorous screening. Subsequently, using NGS sequencing technology, 5 to 15 adapter sequences targeting the target molecule can be obtained, and aptamer binding assays can be performed based on the obtained sequences. Based on its mature SELEX technology platform, the screened nucleic acid aptamers exhibited extraordinary binding affinity, ranging from nM to pM levels. TEK Biotech can provide various nucleotide adapter modifications, including RNA and DNA nucleic acid adapter modification services, by introducing chemical groups or modifiers onto nucleic acid adapter molecules. These chemical modifications can include phosphorylation, methylation, glycosylation, etc. At the same time, nucleic acid adapter PEG modification, various fluorescent and quenching groups (including but not limited to Cy5, Cy7, FITC, etc.) can also be added, and TEK Biotech can purify the modified nucleic acid adapter to ensure its purity and quality. TEK Biotech scientists are able to analyze and design reasonable solutions based on specific experimental needs of customers, provide personalized customized services, and ensure that each customer's unique needs are met.

█ Nucleic Acid Aptamer Modification Service:

The aptamer structure can be folded to form a defined three-dimensional structure and bind to the target protein, thereby inhibiting protein-protein interactions and achieving therapeutic or diagnostic purposes. Aptamers are selected through SELEX technology, and their specificity and affinity are usually equivalent to or better than antibodies.Through SELEX technology, oligonucleotides in the library are repeatedly bound, separated, and amplified with target molecules, gradually screening for aptamers with high affinity and selectivity towards the target molecules. The core of SELEX technology includes nucleic acid molecule libraries, screening schemes, and separation methods between libraries and target molecules. The characteristics of aptamer structure enable it to bind with multiple target molecules with high affinity and selectivity.

Aptamer conjugates are complexes formed by coupling aptamers with small molecule drugs, biotherapies, protein drugs, or nanocarriers. In aptamer conjugates, aptamers act as targets to guide the delivery of other drugs to the target tissue, achieving precise drug delivery and efficient treatment. The aptamer design coupled with small molecules mainly adopts three forms: covalent coupling, nucleic acid synthesis, and physical interaction.

Aptamers have the characteristics of low molecular weight, excellent stability, and ease of chemical synthesis and modification. The core intention of modifying nucleic acid aptamers is to enhance their binding affinity and specificity towards target molecules, thereby optimizing their efficacy. Through chemical modification, the aptamer structure and the charge characteristics of the aptamer can be adjusted, thereby affecting its binding mode and affinity with the target molecule. Moreover, this chemical modification can enhance the stability of the aptamer structure and prolong its half-life in biological systems. According to customer requirements, after synthesizing nucleic acid aptamers with specific sequences and structures, specific chemical groups or modifying components are introduced into the molecular structure of the nucleic acid aptamer, aiming to transform its original properties and functions. These nucleotide aptamer modifications may include various forms such as nucleic acid aptamer PEG modification, phosphorylation, methylation, glycosylation, etc. To ensure that the modified nucleic acid aptamer has high purity and quality, it needs to be purified. Meanwhile, with the help of experimental techniques such as BLI and SPR, the modified nucleic acid aptamers can be validated to confirm whether their binding affinity and specificity meet the standards. Using modified nucleic acid aptamers as recognition components for biosensors can achieve rapid and sensitive detection of specific target molecules. Combining modified nucleic acid aptamers with drug molecules to form aptamer conjugates and targeted drug delivery systems, achieving precise drug delivery and release. The process of TEK Biotech ligand screening, synthesis, and modification is shown in Figure 1.

Adaptation screening, synthesis, and modification process diagram-tekbiotech.jpg

Figure 1 Adaptation screening, synthesis, and modification process diagram

█ Service Content and Cycle:

Step

Service Content

Cycle

Step 1: Construction and screening of nucleic acid

aptamer library

(1) Synthesis of fixed and random sequences, in vitro construction of adapter library.

(2) Using biotin labeled target proteins as screening targets.

(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, modification, and affinity

determination of aptamers (optional)

(1) Synthesize aptamers based on sequences and perform single or double terminal modifications.

(2) Affinity determination of adapter and target protein, KD determination by BLI or SPR.

(3) Delivery: Experimental report, raw data.

6-8 Weeks

█ Service Advantages:


According to the specific experimental needs of customers, there are multiple screening targets to choose from: metal ion, proteins, peptides, amino acids, small molecules, and cells and bacteria, etc	The library has a capacity of 10 ^ 13-10 ^ 14, sufficient to screen for nucleic acid aptamers targeting customer targets	Multiple modification methods are available: DNA nucleic acid aptamer modification, nucleic acid aptamer PEG modification, fluorescent group modification, quenching group modification, etc	SELEX technology platform mature: the affinity of nucleic acid aptamers obtained through screening can reach the nM-pM level

Multiple screening methods: magnetic bead SELEX, cell SELEX, capture SELEX, affinity chromatography SELEX, graphene oxide SELEX, etc. Scientists from Tek Biotech design them reasonably according to specific projects	6-10 rounds of pressure screening can obtain high affinity and high specificity nucleic acid aptamers	Supporting downstream validation experiments: affinity validation (including bli and SPR affinity validation), competitive ELISA validation, flow blocking validation, etc	Widely applicable: The delivered content can be used for molecular interaction analysis of proteins, nucleic acids, peptides, nanomaterials, and other molecules

Nucleic Acid Aptamer Modification Service Frequently Asked Questions

What is the aptamer modification service?

Nucleic aptamer (Aptamer) is a synthetic single-stranded DNA or RNA. Due to its unique tertiary structure, it can bind closely to target molecules and has the characteristics of easy screening, synthesis, and chemical modification. However, because of the large number of reaction sites in the recognition domain, the site-specific chemical modification of the aptamer remains a challenging challenge. Aptamers increase their function or detection effects by introducing various functional groups or markers on their fixed or random sequences. For example, fluorescein, biotin, magnetic beads, and gold nanoparticles can be modified on nucleic acid aptamers for signal amplification or signal transduction; polyethylene glycol (PEG) and cholesterol can be modified on nucleic acid aptamers to increase their solubility or cycle life; drugs and drug-loaded nanoparticles can be modified on nucleic acid aptamers for targeted therapy or controlled release. Modifying aptamers is a significant technology that can improve the stability, specificity, and affinity of aptamers to expand their application in biomedical research. The development of this technology brings new possibilities to the field of genetic engineering and will provide more opportunities and hope for areas such as disease diagnosis and treatment.TEK Biotech can provide a variety of different aptamer modification services, including chemical group modification, PEG modification, protein modification, and so on.
What are the methods of nucleic aptamer modification?

Aptamers include chemical modification, nanomodification, protein modification, and crosslinker modification. Chemical modification is a method to change the original properties of the nucleic acid aptamer by introducing a chemical group at a specific position of the nucleic acid aptamer. These chemical modifications can occur at the base, the sugar ring, or the phosphate backbone of the aptamer. Nanomaterials modification is a method to combine nucleic acid aptamers with nanoparticles such as our common gold nanoparticles and carbon nanotubes to form composite materials. This modification can exploit the unique properties of the nanomaterials themselves to enhance the properties of our desired aptamers. Protein or polypeptide modification is methods that bind aptamers to proteins or peptides, enabling them to form a complex. This modification may enable new biological functions for the aptamer. Coupled modification is a method of coupling the aptamer with a vector or other molecules. This method can help the aptamer to fix or increase the function of the aptamer. After various modifications, the nucleic acid aptamer can improve the original stability, affinity, and specificity of the aptamer, and help the aptamer to play a better function.TEK Biotech can provide one-stop aptamer screening, aptamer synthesis, and aptamer modification services, committed to better-serving customers.
What are the advantages of the separate aptamer modification method?

The modification methods of nucleic aptamers are varied, each with its unique advantages. Chemical modification can enhance the stability of the aptamer under various environmental conditions, such as high temperature, and low pH, which can prolong the half-life of the aptamer in vivo. At the same time, certain chemical modifications can optimize the three-dimensional structure of the aptamer, improve the binding force with the target molecule, and enhance the affinity. Through chemical modification, it can reduce the rejection reaction triggered by the aptamer after entering the body and reduce its immune properties. Because of the good electrical conductivity, the nanoaptamer can improve the sensitivity of the nanoaptamer, so that the aptamer can reach the target position more accurately, and achieve accurate treatment or diagnosis. Modification of protein or polypeptide can increase the biological activity of the aptamer, enabling it to play better effects in the body, and at the same time, the protein can prolong the circulation time of the aptamer in the body, and cooperate with the aptamer to improve the overall targeting effect. The method of coupling aptamers is relatively simple and convenient for experimenters to operate and control. Through coupling, the aptamer can be combined with various functional molecules to realize the multifunctional application of the aptamer.
How to maintain the serum stability of the selected aptamer?

In recent years, the focus on aptamers has increased, and the development of therapeutic drugs has become a focus of current research. However, as a member of oligonucleotides, aptamer also has the characteristics of oligonucleotides, which are easy to be degraded by nucleases, which will affect the stability and function of aptamer in serum. To address these questions, the scientists came up with modifying the aptamers to prevent the degradation of the nucleases. Nonetheless, studies on how to improve aptamer stability in serum by modifying the aptamers are still lacking. There has been a recent study of interest that systematically evaluated several chemically modified oligonucleotide sequences, and stability performance in frozen human serum, freshly collected mouse serum, and freshly collected human serum. The researchers found that adding a cap containing an inverted dT before the sequence gently enhanced serum stability and that the modified aptamer stability was instead increased compared to the first variant. In contrast, those aptamers are better stable, have the longest serum half-life, and have no particularly obvious degradation even after prolonged exposure to organisms. This study provides a more stable and reliable basis for the future development and application of aptamer drugs.TEK Biotech improves modified aptamer services to help improve the stability of aptamers in serum.
The efficiency of the aptamer modification and the modification specificity problem, and its solution?

When we add chemical groups to the nucleic acid aptamers for some modifications, we may find that the modification efficiency of the chemical groups is not high, resulting in the waste of the synthetic aptamers and a low yield after modification. Faced with the problem of post-modification efficiency, we helped improve the modification efficiency of chemical groups or other substances, such as optimizing the modification reaction conditions, such as the reaction temperature, the pH, the reaction time, and the concentration of the modifier. At the same time, we also need to ensure the purity of the whole reaction system and reduce the interference of impurities in the system. In addition to the low modification efficiency, the modified groups may not appear in the expected site and appear in other positions, which will reduce the specificity of the nucleic acid aptamer. Therefore, when we choose the modification, we need to analyze the modified sequence to ensure the accuracy of the final results. TEK Biotech has been deeply engaged in the nucleic acid aptamer modification platform for many years, using a rich screening and modification platform and excellent technical services to solve problems for customers in one station and help customers in scientific research, drug development, etc.

What is the aptamer modification service?

Nucleic aptamer (Aptamer) is a synthetic single-stranded DNA or RNA. Due to its unique tertiary structure, it can bind closely to target molecules and has the characteristics of easy screening, synthesis, and chemical modification. However, because of the large number of reaction sites in the recognition domain, the site-specific chemical modification of the aptamer remains a challenging challenge. Aptamers increase their function or detection effects by introducing various functional groups or markers on their fixed or random sequences. For example, fluorescein, biotin, magnetic beads, and gold nanoparticles can be modified on nucleic acid aptamers for signal amplification or signal transduction; polyethylene glycol (PEG) and cholesterol can be modified on nucleic acid aptamers to increase their solubility or cycle life; drugs and drug-loaded nanoparticles can be modified on nucleic acid aptamers for targeted therapy or controlled release. Modifying aptamers is a significant technology that can improve the stability, specificity, and affinity of aptamers to expand their application in biomedical research. The development of this technology brings new possibilities to the field of genetic engineering and will provide more opportunities and hope for areas such as disease diagnosis and treatment.TEK Biotech can provide a variety of different aptamer modification services, including chemical group modification, PEG modification, protein modification, and so on.
What are the methods of nucleic aptamer modification?

Aptamers include chemical modification, nanomodification, protein modification, and crosslinker modification. Chemical modification is a method to change the original properties of the nucleic acid aptamer by introducing a chemical group at a specific position of the nucleic acid aptamer. These chemical modifications can occur at the base, the sugar ring, or the phosphate backbone of the aptamer. Nanomaterials modification is a method to combine nucleic acid aptamers with nanoparticles such as our common gold nanoparticles and carbon nanotubes to form composite materials. This modification can exploit the unique properties of the nanomaterials themselves to enhance the properties of our desired aptamers. Protein or polypeptide modification is methods that bind aptamers to proteins or peptides, enabling them to form a complex. This modification may enable new biological functions for the aptamer. Coupled modification is a method of coupling the aptamer with a vector or other molecules. This method can help the aptamer to fix or increase the function of the aptamer. After various modifications, the nucleic acid aptamer can improve the original stability, affinity, and specificity of the aptamer, and help the aptamer to play a better function.TEK Biotech can provide one-stop aptamer screening, aptamer synthesis, and aptamer modification services, committed to better-serving customers.
What are the advantages of the separate aptamer modification method?

The modification methods of nucleic aptamers are varied, each with its unique advantages. Chemical modification can enhance the stability of the aptamer under various environmental conditions, such as high temperature, and low pH, which can prolong the half-life of the aptamer in vivo. At the same time, certain chemical modifications can optimize the three-dimensional structure of the aptamer, improve the binding force with the target molecule, and enhance the affinity. Through chemical modification, it can reduce the rejection reaction triggered by the aptamer after entering the body and reduce its immune properties. Because of the good electrical conductivity, the nanoaptamer can improve the sensitivity of the nanoaptamer, so that the aptamer can reach the target position more accurately, and achieve accurate treatment or diagnosis. Modification of protein or polypeptide can increase the biological activity of the aptamer, enabling it to play better effects in the body, and at the same time, the protein can prolong the circulation time of the aptamer in the body, and cooperate with the aptamer to improve the overall targeting effect. The method of coupling aptamers is relatively simple and convenient for experimenters to operate and control. Through coupling, the aptamer can be combined with various functional molecules to realize the multifunctional application of the aptamer.
How to maintain the serum stability of the selected aptamer?

In recent years, the focus on aptamers has increased, and the development of therapeutic drugs has become a focus of current research. However, as a member of oligonucleotides, aptamer also has the characteristics of oligonucleotides, which are easy to be degraded by nucleases, which will affect the stability and function of aptamer in serum. To address these questions, the scientists came up with modifying the aptamers to prevent the degradation of the nucleases. Nonetheless, studies on how to improve aptamer stability in serum by modifying the aptamers are still lacking. There has been a recent study of interest that systematically evaluated several chemically modified oligonucleotide sequences, and stability performance in frozen human serum, freshly collected mouse serum, and freshly collected human serum. The researchers found that adding a cap containing an inverted dT before the sequence gently enhanced serum stability and that the modified aptamer stability was instead increased compared to the first variant. In contrast, those aptamers are better stable, have the longest serum half-life, and have no particularly obvious degradation even after prolonged exposure to organisms. This study provides a more stable and reliable basis for the future development and application of aptamer drugs.TEK Biotech improves modified aptamer services to help improve the stability of aptamers in serum.
The efficiency of the aptamer modification and the modification specificity problem, and its solution?

When we add chemical groups to the nucleic acid aptamers for some modifications, we may find that the modification efficiency of the chemical groups is not high, resulting in the waste of the synthetic aptamers and a low yield after modification. Faced with the problem of post-modification efficiency, we helped improve the modification efficiency of chemical groups or other substances, such as optimizing the modification reaction conditions, such as the reaction temperature, the pH, the reaction time, and the concentration of the modifier. At the same time, we also need to ensure the purity of the whole reaction system and reduce the interference of impurities in the system. In addition to the low modification efficiency, the modified groups may not appear in the expected site and appear in other positions, which will reduce the specificity of the nucleic acid aptamer. Therefore, when we choose the modification, we need to analyze the modified sequence to ensure the accuracy of the final results. TEK Biotech has been deeply engaged in the nucleic acid aptamer modification platform for many years, using a rich screening and modification platform and excellent technical services to solve problems for customers in one station and help customers in scientific research, drug development, etc.

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