title: 3,3'-Diamino-N-methyldipropylamine as a versatile affinity ligand.
abstract: Currently, in biomedicine and biotechnology fields, there is a growing need to develop and produce biomolecules with a high degree of purity. To accomplish this goal, new purification methods are being developed looking for higher performance, efficiency, selectivity, and cost-effectiveness. Affinity chromatography is considered one of the most highly selective methods for biomolecules purification. The purpose of this work is to explore a new type of a structurally simple ligand immobilized onto an agarose matrix to be used in affinity chromatography. The ligand in this study, 3,3'-diamino-N-methyldipropylamine has shown low toxicity and low cost of preparation. Moreover, the ability of the ligand to be used in affinity chromatography to purify proteins and nucleic acids was verified. An increasing sodium chloride gradient, using salt concentrations up to 500 mM, was suitable to accomplish the purification of these biomolecules, meaning that the new support allows the recovery of target biomolecules under mild conditions. Thus, the 3,3'-diamino-N-methyldipropylamine ligand is shown to be a useful and versatile tool in chromatographic experiments, with very good results either for proteins or supercoiled plasmid isoform purification.
authors.affiliation: CICS-UBI-Centro de Investigação em Ciências da Saúde, Universidade da Beira Interior, Covilhã, Portugal.
authors.forename: Marta S
authors.initials: MS
authors.lastname: Silva
authors.forename: Vânia C
authors.initials: VC
authors.lastname: Graça
authors.forename: Lucinda V
authors.initials: LV
authors.lastname: Reis
authors.forename: Paulo F
authors.initials: PF
authors.lastname: Santos
authors.forename: Samuel
authors.initials: S
authors.lastname: Silvestre
authors.forename: Luiza
authors.initials: L
authors.lastname: Granadeiro
authors.forename: Paulo
authors.initials: P
authors.lastname: Almeida
authors.forename: João A
authors.initials: JA
authors.lastname: Queiroz
authors.forename: Fani
authors.initials: F
authors.lastname: Sousa
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D004278:DNA, Superhelical / Q000737:chemistry / Q000302:isolation & purification*; D008024:Ligands; D010957:Plasmids / Q000737:chemistry / Q000302:isolation & purification*; D011073:Polyamines / Q000737:chemistry*; D011437:Propylamines / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012685:Sepharose / Q000737:chemistry
chemical_list: D004278:DNA, Superhelical; D008024:Ligands; D011073:Polyamines; D011437:Propylamines; D011506:Proteins; C000602661:3,3'-diamino-N-methyldipropylamine; D012685:Sepharose
keywords: 3,3′-Diamino-N-methyldipropylamine ligand; Affinity chromatography; Supercoiled plasmid DNA