A kind of ortho-phthalaldehyde derivative phosphoramidite monomer, its synthesis method and the method for dna rapid coupling protein

Abstract

The invention relates to an o-phthaldehyde derivative phosphoramidite monomer, a synthesizing method and a method for quick coupling of DNA (deoxyribonucleic acid) and protein, and relates to quick coupling of DNA and protein. A molecular formula of the o-phthaldehyde derivative phosphoramidite monomer is C28H46N3O6P. The synthesizing method comprises the following steps of firstly, enabling o-phthaldehyde carboxylic acid derivative (OPA-COOH) and 6-amino-1-hexanol to react, so as to prepare the o-phthaldehyde hydroxyl derivative (OPA-OH); then, under the conditions of nitrogen protection, a dichloromethane solvent and N,N-diisopropylethylamine, reacting with 2-cyanoethyl-N,N-diisopropylchlorophosphoramidite for 2h at room temperature; separating by silica gel column chromatography, so as to obtain the o-phthaldehyde derivative phosphoramidite monomer. The reaction characteristic of adjacent aldehyde groups of the o-phthaldehyde derivative and the amino groups on the protein lysine residue groups quickly and efficiently generating phthalimidine is utilized, so that the purpose of quick coupling of OPA-DNA and natural protein is realized. Compared with other coupling methods, the method has the advantages that the selectivity is realized, the operation is quick, and the efficiency is high.

Description

technical field

[0001] The invention relates to a phthalaldehyde derivative phosphoramidite monomer and a synthesis method thereof, in particular to a method for fast coupling of DNA and protein. Background technique

[0002] The modification of natural proteins is of great significance to the field of bioanalysis research. For example, in biosensors, the coupling of DNA and proteins is usually required. Traditional DNA-protein coupling methods are mainly divided into two categories, namely non-covalent coupling and covalent coupling. The most common non-covalent coupling method is through the non-covalent binding of biotin and streptavidin. Due to their very strong non-covalent binding ability, they are widely used in the field of biological analysis, such as enzyme-linked immunosorbent adsorption. Assay (ELISA). However, the above-mentioned coupling system is cumbersome, requires the help of streptavidin with a relatively large molecular weight, and the modification ste...

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