Apta-Index™: Human Neutrophil Elastase (DNA I) (ID# 175)

Human Neutrophil Elastase (DNA I) (ID# 175)

* The secondary structure may not be accurate because of MFOLD limitations on chemistries other than RNA/DNA.

Aptamer Chemistry: DNA Target: Human Neutrophil Elastase (HNE) Antigen/Target Category: Protein Affinity (Kd): 17 nM (reported value) Binding Conditions/Buffer: Binding buffer: 150 mM NaCl, 100 mM Tris HCl (pH 7.0), 2 mM MgCl₂, 6 mM KCl Refolding Program: N/A If the oligo is a known aptamer sequence: For binding studies, perform a refolding program to ensure proper function (i.e. binding to antigen or target). Refer to the aptamer reference source for the appropriate refolding parameters and binding conditions. Note: it is unknown whether aptamer functions properly without refolding. Comments: This truncated sequence forms a G-quartet.

Neither the full-length, nor the truncated sequence inhibited HNE enzymatic activity, as the aptamer binding site differs from the active site and the substrate binding site.

Sequence (): 5'-dTpdApdGpdCpdGpdApdTpdApdCpdTpdGpdCpdGpdTpdGpdGpdGpdTpdTpdGpdGpdGpdGpdCpdGpdGpdGpdTpdApdGpdGpdGpdCpdCpdApdGpdCpdApdGpdTpdCpdTpdCpdGpdTp-3'
Length: 45 Molecular Weight: 14046.12 g/mole Extinction Coefficient: 428800.00 L/(mole·cm) GC Content: 64.44% nmoles/OD₂₆₀: 2.33 μg/OD₂₆₀: 32.76

Note: Information on this aptamer oligo was obtained from the literature and hasn't been validated by Aptagen.

Reference:

Lin et al. "Peptide conjugation to an in vitro-selected DNA ligand improves enzyme inhibition." PNAS, 92(1995): 11044-11048.

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Aptagen develops and manufactures aptamers (synthetic antibodies) which are ligands of RNA, DNA, and peptide oligos that bind to a variety of target antigens. Aptamers (synthetic antibodies) are sometimes referred to as "chemical antibodies or DNA antibodies." Aptazymes are aptamers (synthetic antibodies) with enzymatic activity. A subclass of aptazymes called molecular switches is analogous to molecular beacons for fluorescent detection of analytes in test samples. Examples include allosteric ribozymes, also known as riboswitches.

Allosteric ribozymes (or riboswitches) have been engineered to act like molecular switches turning "on" or "off" in the presence or absence of effectors. Ribozyme catalysis is modulated by rapid conformational changes imparted by an effector molecule binding to an aptamer domain apart from the catalytic site. The aptamer domain is a structural region of several nucleotides that binds specifically to its cognate effector molecule. With the advent of allosteric RNA switches and aptazymes, it was soon recognized that the precise molecular recognition and high degree of selectively exhibited by these molecules make them ideal biosensor elements which can be immobilized on a solid support for microarray analysis and biosensor applications. Compared to other types of biosensor recognition elements such as proteins, antibodies, and oligonucleotides, which are commonly used in microarrays to detect a narrow assortment of targets, RNA allosteric elements are better suited for the detection of a wide variety of analytes in a complex mixture, since unlike proteins, antibodies, and oligonucleotides, they are not limited by the types of targets they can bind. As described before, the ability to rapidly develop RNA molecular switches using one pot selection techniques against a vast array of targets permits the rapid development of biochips capable of analyzing and fingerprinting a complex mixture of chemical and biological targets. The versatility and utility of these molecular switches have been employed in biochips which are able to detect specific targets from a complex mixture containing metal ions, enzyme cofactors, metabolites, serum biomarkers, drug analytes, and strains of E coli strain from culture media.

Aptagen, LLC is a biotechnology company offering aptamer (synthetic antibody) products and services as research reagents, diagnostic and biomarker discovery tools, as well as for use in drug discovery and targeted delivery for therapeutics, and bioindustrial applications. We have 20 years of experience in developing aptamers (synthetic antibodies) for all types of downstream applications.

Apta-Index™: Human Neutrophil Elastase (DNA I) (ID# 175)

Human Neutrophil Elastase (DNA I) (ID# 175)

Lin et al. "Peptide conjugation to an in vitro-selected DNA ligand improves enzyme inhibition." PNAS, 92(1995): 11044-11048.

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