title: Deprotection Reagents in Fmoc Solid Phase Peptide Synthesis: Moving Away from Piperidine?
abstract: The deprotection step is crucial in order to secure a good quality product in Fmoc solid phase peptide synthesis. 9-Fluorenylmethoxycarbonyl (Fmoc) removal is achieved by a two-step mechanism reaction favored by the use of cyclic secondary amines; however, the efficiency of the reaction could be affected by side reactions and by-product formation. Several aspects have to be taken into consideration when selecting a deprotection reagent: its physicochemical behavior, basicity (pKa) and polarity, concentration, and time of reaction, toxicity and disposability of residues and, finally, availability of reagents. This report presents a comparison of the performance of three strategies for deprotection using microwave-assisted Fmoc peptide synthesis. Four peptide sequences were synthesized using Rink amide resin with a Liberty Blue™ automated synthesizer and 4-methylpiperidine (4MP), piperidine (PP), and piperazine (PZ) as Fmoc removal reagents. In the first instance all three reagents behaved similarly. A detailed analysis showed a correlation between the hydrophobicity and size of the peptide with the yield and purity of the obtained product. The three reagents are interchangeable, and replacement of piperidine could be advantageous regarding toxicity and reagent handling.
authors.affiliation: Fraunhofer Chile Research, Santiago 7550296, Chile. omar.luna@fraunhofer.cl.
authors.forename: Omar F
authors.initials: OF
authors.lastname: Luna
authors.affiliation: Núcleo Biotecnología Curauma, Pontificia Universidad Católica de Valparaíso, Valparaíso 2373223, Chile. johanagomezuribe@gmail.com.
authors.forename: Johana
authors.initials: J
authors.lastname: Gomez
authors.affiliation: Fraunhofer Chile Research, Santiago 7550296, Chile. constanza.cardenas@pucv.cl.|Núcleo Biotecnología Curauma, Pontificia Universidad Católica de Valparaíso, Valparaíso 2373223, Chile. constanza.cardenas@pucv.cl.
authors.forename: Constanza
authors.initials: C
authors.lastname: Cárdenas
authors.affiliation: CIBER-BBN, Networking Centre on Bioengineering, Biomaterials and Nanomedicine, University of Barcelona, Barcelona 08028, Spain. albericio@ub.edu.|Department of Organic Chemistry, University of Barcelona, Barcelona 08028, Spain. albericio@ub.edu.|School of Chemistry, University of KwaZulu-Natal, Durban 4001, South Africa. albericio@ub.edu.
authors.forename: Fernando
authors.initials: F
authors.lastname: Albericio
authors.affiliation: Fraunhofer Chile Research, Santiago 7550296, Chile. sergio.marshall@pucv.cl.|Núcleo Biotecnología Curauma, Pontificia Universidad Católica de Valparaíso, Valparaíso 2373223, Chile. sergio.marshall@pucv.cl.|Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Valparaíso 2373223, Chile. sergio.marshall@pucv.cl.
authors.forename: Sergio H
authors.initials: SH
authors.lastname: Marshall
authors.affiliation: Fraunhofer Chile Research, Santiago 7550296, Chile. fanny.guzman@pucv.cl.|Núcleo Biotecnología Curauma, Pontificia Universidad Católica de Valparaíso, Valparaíso 2373223, Chile. fanny.guzman@pucv.cl.
authors.forename: Fanny
authors.initials: F
authors.lastname: Guzmán
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D005449:Fluorenes / Q000737:chemistry; D007700:Kinetics; D010455:Peptides / Q000138:chemical synthesis*; D010880:Piperidines / Q000737:chemistry; D060327:Solid-Phase Synthesis Techniques; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000596:Amino Acids; D005449:Fluorenes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D010455:Peptides; D010880:Piperidines
keywords: 4-methylpiperidine; N-α-deprotection reagent; piperazine; piperidine; solid-phase peptide synthesis