Support For Solid-phase Peptide Synthesis Research Articles

Acid-Modulated Peptide Synthesis for Application on Oxide Biosensor Interfaces.

In this paper we report an acid-modulated strategy for novel peptide microarray production on biosensor interfaces. We initially selected a controlled pore glass (CPG) as a support for solid-phase peptide synthesis (SPPS) to implement a chemistry that can be performed at the interface of multiple field effect transistor (FET) sensors, eventually to generate label-free peptide microarrays for protein screening. Our chemistry uses a temporary protection of the N-terminal amino function of each amino acid building block with a tert-butyloxycarbonyl (Boc) group that can be removed after each SPPS cycle, in combination with semi-permanent protection of the side chains of trifunctional amino acid residues. Such a protection scheme with a well-proven record of application in conventional, batchwise SPPS has been fine-tuned for optimal performance on CPG and, from there, translated to SPR chips that allow layer-by-layer monitoring of amino acid coupling. Our results validate this acid-modulated synthesis as a feasible approach for producing peptides in high yields and purity on flat glass surfaces, such as those in bio-FETs.

OctaGel Resin - A New PEG-PS-based Solid Support for Solid-Phase Peptide Synthesis

: OctaGel resin is a unique, highly uniformed surface-active resin. Here, we compared the performance of OctaGel with that of known resins on the market, namely polystyrene and ChemMatrix, in Solid-Phase Peptide Synthesis. The synthesis of the ‘difficult’ Aib-ACP (65-74) decapeptide showed that OctaGel has the potential to yield molecules with satisfactory purity. Given its high swelling capacity and large bead size, OctaGel also shows efficient interaction with various solvents, including those mainly used for SPPS (DMF and DCM).

Preparation of tri(ethylene glycol) grafted core-shell type polymer support for solid-phase peptide synthesis.

A core-shell type polymer support for solid-phase peptide synthesis has been developed for high coupling efficiency of peptides and versatile applications such as on-bead bioassays. Although various kinds of polymer supports have been developed, they have their own drawbacks including poor accessibility of reagents and incompatibility in aqueous solution. In this paper, we prepared hydrophilic tri(ethylene glycol) (TEG) grafted core-shell type polymer supports (TEG SURE) for efficient solid-phase peptide synthesis and on-bead bioassays. TEG SURE was prepared by grafting TEG derivative on the surface of AM PS resin via biphasic diffusion control method and subsequent acetylation of amine groups which are located at the core region of AM PS resin. The performance of TEG SURE was evaluated by synthesizing several peptides. Three points can be highlighted: (1) easy control of loading level of TEG, (2) improved efficiency of peptide synthesis compared with the conventional resins, and (3) applicability of on-bead bioassays.

Multifunctional ionic liquid-bound polystyrene resin with high loading capacity as support in solid-phase peptide synthesis

Polystyrene resin-bound ionic liquids (PSILs) with high loading capacities were prepared by immobilizing multifunctional ionic liquids (ILs) on modified polystyrene (PS) resin and used in the solid phase peptide synthesis. Introduction of hydrophobic anions and functional side chain containing ILs resulted in high yield (82–98%) and purity (92–98%) of the synthesized peptides. The coupling kinetic studies of the first and second amino acids to the PSILs were performed to investigate the effect of IL functionalization on PS supports.

Multilayered Magnetic Nanoparticles as a Support in Solid-Phase Peptide Synthesis

The synthesis of multilayered magnetic nanoparticles (MNPs) for use as a support in solid-phase peptide synthesis (SPPS) is described. Silanization of magnetite (Fe3O4) nanoparticles with 3-(trimethoxysilyl)propyl methacrylate introduced polymerizable groups on the surface. Polymerization with allylamine, trimethylolpropane trimethacrylate, and trimethylolpropane ethoxylate (14/3 EO/OH) triacrylate provided a polymeric coating and amino groups to serve as starting points for the synthesis. After coupling of an internal reference amino acid and a cleavable linker, the coated MNPs were applied as the solid phase during synthesis of Leu-enkephalinamide and acyl carrier protein (65-74) by Fmoc chemistry. A “high-load” version of the MNP support (0.32 mmol/g) was prepared by four consecutive cycles of Fmoc-Lys(Fmoc)-OH coupling and Fmoc deprotection. Successful synthesis of Leu-enkephalin was demonstrated on the “high-load” MNPs. Chemical stability studies proved the particles to be stable under SPPS conditions and magnetization measurements showed that the magnetic properties of the particles were maintained throughout derivatizations and SPPS. The MNPs were further characterized by high-resolution transmission electron microscopy, inductively coupled plasma atomic emission spectrometry, elemental analysis, and nitrogen gas adsorption measurements.

A new method for the preparation of 2-chlorotrityl resin and its application to solid-phase peptide synthesis

2-Chlorotritylchloride (2-CTC) resin was prepared efficiently from 1% DVB-crosslinked polystyrene resin and 1-chloro-2-(dichloro(phenyl)methyl)benzene, which was easily obtained from 2-chlorobenzophenone. This 2-CTC resin showed excellent properties as a support for solid-phase peptide synthesis. Four peptide fragments were obtained in high purity using the resin.

A novel chemically and mechanically stable glycerol-based crosslinked polystyrene support for polypeptide synthesis: A comparative study with Merrifield resin

A novel chemically and mechanically stable polymer support for solid-phase peptide synthesis with excellent swelling performance in various organic solvents is described. The crosslinked polymer in its bead form was prepared by the aqueous suspension polymerization of the styrene and tri(propylene glycol) glycerolate diacrylate. The support is unique because the hydroxyl group is introduced into the polymer support in the polymerization step itself, which makes it highly cost effective. The hydroxyl functionality of the crosslinker in the polymer was used as a growth site for peptide synthesis, thus minimizing the incompatibility arising because of the styrene core as compared to styrene-based polymer supports currently used in polypeptide synthesis. The utility of the resin for solid-phase peptide synthesis was tested by the synthesis of a 19-residue peptide amide of the envelope region of hepatitis C viral polyprotein and was compared with Merrifield resin by the 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl uronium hexafluoro phosphate/1-hydroxy benzatriazole method with a Rink amide linker. The molecular character of the polymer and the extent of crosslinking density on the dependence of the reactivity of the attached amino groups were investigated by a reactivity study of the amide bond formation of a model Val-Ala dipeptide. Enhanced swelling, the increased rate of aminoacylation, and the high purity of the peptides synthesized on the novel support highlighted the influence of the polarity of the solid support in the solid-phase synthesis. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 92: 288–294, 2004

SUBPOL: a novel SUcrose-Based Polymer support for solid-phase peptide synthesis and affinity chromatography applications.

A novel SUcrose-Based Polymer support (SUBPOL) with tailored morphology suitable for the use in solid-phase peptide synthesis (SPPS) is described, and its application as a hydrophilic affinity matrix for the specific removal of fibrinogen from human plasma is demonstrated. After suspension polymerization of partly methacrylated 2,1':4,6-di-O-isopropylidene sucrose and subsequent removal of the protecting groups, hydrophilic spherical polymer beads were obtained. The morphology of the resulting resin was controlled by variation of the porogen as well as the average degree of substitution with respect to the methacryloyl groups of the monomer mixture. After introduction of amino groups for a permanent attachment of immobilized peptide ligands, prevention of unintended esterification during SPPS was achieved by silylation of remaining hydroxy groups. Alternatively, a Rink amide linker was introduced prior to SPPS to allow cleavage and subsequent analysis of the peptide assembled on the SUBPOL resins. Two hexapeptides of sequence kwiivw and hffflw, consisting of d-amino acids, as well as a 19-mer peptide corresponding to the sequence GSGVRGDFGSLAPRVARQL of the VP1 protein from the foot-and-mouse disease virus (FMDV) were successfully prepared both manually or in a semi-automated process on SUBPOL resins according to the Fmoc/tBu strategy. Yields and purities were comparable to peptides prepared on commercially available polystyrene resins. A specific affinity adsorbent containing the fibrinogen-binding pentapeptide GPRPK was prepared by SPPS on SUBPOL resins of different morphology, and the strong impact of the affinity matrix on adsorption performance was demonstrated.

Side-chain anchoring strategy for solid-phase synthesis of peptide acids with C-terminal cysteine.

Many naturally occurring peptide acids, e.g., somatostatins, conotoxins, and defensins, contain a cysteine residue at the C-terminus. Furthermore, installation of C-terminal cysteine onto epitopic peptide sequences as a preliminary to conjugating such structures to carrier proteins is a valuable tactic for antibody preparation. Anchoring of N(alpha)-Fmoc, S-protected C-terminal cysteine as an ester onto the support for solid-phase peptide synthesis is known to sometimes occur in low yields, has attendant risks of racemization, and may also result in conversion to a C-terminal 3-(1-piperidinyl)alanine residue as the peptide chain grows by Fmoc chemistry. These problems are documented for several current strategies, but can be circumvented by the title anchoring strategy, which features the following: (a). conversion of the eventual C-terminal cysteine residue, with Fmoc for N(alpha)-amino protection and tert-butyl for C(alpha)-carboxyl protection, to a corresponding S-xanthenyl ((2)XAL(4)) preformed handle derivative; and (b). attachment of the resultant preformed handle to amino-containing supports. This approach uses key intermediates that are similar to previously reported Fmoc-XAL handles, and builds on earlier experience with Xan and related protection for cysteine. Implementation of this strategy is documented here with syntheses of three small model peptides, as well as the tetradecapeptide somatostatin. Anchoring occurs without racemization, and the absence of 3-(1-piperidinyl)alanine formation is inferred by retention of chains on the support throughout the cycles of Fmoc chemistry. Fully deprotected peptides, including free sulfhydryl peptides, are released from the support in excellent yield by using cocktails containing a high concentration (i.e., 80-90%) of TFA plus appropriate thiols or silanes as scavengers. High-yield release of partially protected peptides is achieved by treatment with cocktails containing a low concentration (i.e., 1-5%) of TFA. In peptides with two cysteine residues, the corresponding intramolecular disulfide-bridged peptide is obtained by either (a). oxidation, in solution, of the dithiol product released by acid; (b). simultaneous acidolytic cleavage and disulfide formation, achieved by addition of the mild oxidant DMSO to the cleavage cocktail; or (c). concomitant cleavage/cooxidation (involving a downstream S-Xan protected cysteine), using reagents such as iodine or thallium tris(trifluoroacetate) in acetic acid.

A comparison of rigid and flexible crosslinked polymer-supported peptide synthesis

To correlate the coupling difficulty and nature of the support in solid-phase peptide synthesis, a set of difficult peptides were built under identical conditions on rigid and hydrophobic divinyl benzene-crosslinked polystyrene (DVB–PS) and flexible and polar 1,4-butanediol dimethacrylate-crosslinked polystyrene (BDDMA–PS) resins of same functional capacity (∼2 mmol/g). The difficult coupling stages observed on each support were identified by monitoring the acylation time, coupling yield and deprotection efficiency. The yield and purity of the peptides prepared on BDDMA–PS were higher than when DVB–PS support was used.

A novel core-shell type polymer support for solid-phase peptide synthesis

Novel core-shell type resins with a highly cross-linked polystyrene (PS) core and a poly(ethylene glycol) (PEG) shell were prepared by suspension polymerization for use in solid-phase synthesis. All the resins were of bead type with a diameter of 50–200 μm. The loading capacities of the amino groups were 0.05–0.2 mmol/g resin and the thickness of the PEG shell was 2–4 μm. Compared with the other resins such as 1% cross-linked PS resin and TentaGel resin, the resins showed excellent performance in photolytic release of peptide product from photolabile linker.

Peptide synthesis on chitin.

The use of chitin as a support for solid-phase peptide synthesis is described and illustrated by synthesis of four peptides, varying in length from 10 to 29 residues. Syntheses were performed in a continuous-flow peptide synthesizer, using Fmoc chemistry. A cleavable linker, p-[(R,S)-alpha-[1-(9H-fluoren-9-yl)-methoxyformamido]-2,4-di methoxybenzyl]- phenoxyacetic acid, was attached to chitosan at the desired substitution level, and the complex acetylated to yield a linker substituted chitin. The effects of temperature, solvents and degree of linker substitution on the syntheses were studied. Acyl carrier peptide (ACP) synthesis studies indicated that temperature was the single most important parameter. Increasing the temperature of the synthesis from 20 to 55 degrees C resulted in an enormous improvement of this synthesis, with about 90% of the crude product being the correct peptide. Denaturing solvents, such as DMSO, could be used without significant effect on the flow properties of the support. The synthesis of one peptide was mainly improved by lowering the degree of substitution from 0.3 to 0.1 mmol/g, suggesting peptide aggregation was a problem in this case. The results of three syntheses on chitin were comparable with those obtained with a commonly used commercial support. This work shows that, under appropriate conditions, chitin can be utilized directly as a support for peptide synthesis.

Gel-phase peptide synthesis on a new high-capacity tetraethyleneglycol diacrylate-crosslinked polystyrene support: Synthesis of pardaxin 16–33.

An insoluble but highly solvating copolymer composed of tetraethyleneglycol diacrylate (TTEGDA)-crosslinked polystyrene has been prepared in beaded form by free-radical suspension polymerization of the monomers and employed as a new solid support for gel-phase peptide synthesis. This new polymer support has comparable physical and mechanical properties as that of the divinylbenzene (DVB)-crosslinked polystyrene support permitting identical manipulation such as shaking and filtration when used as a support for solid-phase peptide synthesis. This polystyrene-TTEGDA crosslinked polystyrene resin undergoes much more effective swelling and solvation than DVB-crosslinked polystyrene in a range of solvents with widely varying polarity which are commonly employed synthesis. The resin can be easily chloromethylated under controlled conditions to prepare resins of varying capacity ranging from low (0.15 meq/g) to high capacity (3.5meq/g). A high capacity resin with 1.5 meq/g chlorine capacity was used for synthesis of hydrophilic C-terminal 18-residue peptide of pardaxin from Pardachirus Pavoninus. The peptide was cleaved from the polymeric support by trifluoroacetic acid at 37°C and purified by Fast Protein Liquid Chromatography (FPLC). The purified peptide was shown to be homogeneous by HPLC and the identity of the 18-residue peptide was confirmed by amino acid analysis. The free peptide possesses a disordered conformation as revealed by the CD measurement.

Long-chain polystyrene-grafted polyethylene film matrix: a new support for solid-phase peptide synthesis

Long-chain polystyrene-grafted polyethylene film matrix: a new support for solid-phase peptide synthesis

Solid-phase synthesis using a new polyacrylic resin : Synthesis of the fragment 14–21 of the amino acid sequence of histone H4

The solid-phase synthesis of the octapeptide 1 AcGly-Ala-Lys-Arg-His-Arg-Lys-ValOMe, which represents the fragment 14-21 of the amino acid sequence of the chromosomal histone H4, as well as of the structurally related nonapeptide 2 AcGly-Ala-Lys-Leu-Arg-His-Arg-Lys-ValOMe, is described using a new polyacrylic resin containing a glycolamide ester linkage(resin-NHCO-CH 2-OCO-peptide) acting as a labile anchoring moiety between the resin and the peptide. After elongation of the polypeptide chain using classical protecting groups, i.e. t-butyloxycarbonyl for the α-NH 2 function, benzyloxycarbonyl, nitro and 2,4-dinitrophenyl groups for the side-chains of Lys, Arg and His respectively, both peptides 1 and 2 were obtained in good yields and with a high purity as shown by high-pressure liquid chromatography, by amino-acid analysis and by high-field proton NMR spectroscopy. This work demonstrates the ability of the newly introduced polyacrylic resin to act as a convenient support for solid-phase peptide synthesis.

ChemInform Abstract: A NEW SYNTHETIC ROUTE TO TERT‐BUTYLOXYCARBONYLAMINOACYL‐4‐(OXYMETHYL)PHENYLACETAMIDOMETHYL‐RESIN, AN IMPROVED SUPPORT FOR SOLID‐PHASE PEPTIDE SYNTHESIS

Chemischer InformationsdienstVolume 10, Issue 1 Natural Products ChemInform Abstract: A NEW SYNTHETIC ROUTE TO TERT-BUTYLOXYCARBONYLAMINOACYL-4-(OXYMETHYL)PHENYLACETAMIDOMETHYL-RESIN, AN IMPROVED SUPPORT FOR SOLID-PHASE PEPTIDE SYNTHESIS A. R. MITCHELL, A. R. MITCHELLSearch for more papers by this authorS. B. H. KENT, S. B. H. KENTSearch for more papers by this authorM. ENGELHARD, M. ENGELHARDSearch for more papers by this authorR. B. MERRIFIELD, R. B. MERRIFIELDSearch for more papers by this author A. R. MITCHELL, A. R. MITCHELLSearch for more papers by this authorS. B. H. KENT, S. B. H. KENTSearch for more papers by this authorM. ENGELHARD, M. ENGELHARDSearch for more papers by this authorR. B. MERRIFIELD, R. B. MERRIFIELDSearch for more papers by this author First published: January 2, 1979 https://doi.org/10.1002/chin.197901315AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume10, Issue1January 2, 1979 RelatedInformation

A new synthetic route to tert-butyloxycarbonylaminoacyl-4-(oxymethyl)phenylacetamidomethyl-resin, an improved support for solid-phase peptide synthesis

A new synthetic route to tert-butyloxycarbonylaminoacyl-4-(oxymethyl)phenylacetamidomethyl-resin, an improved support for solid-phase peptide synthesis

Tert-butoxycarbonylaminoacyl-4-(oxymethyl)-phenylacetamidomethyl-resin, a more acid-resistant support for solid-phase peptide synthesis.

Tert-butoxycarbonylaminoacyl-4-(oxymethyl)-phenylacetamidomethyl-resin, a more acid-resistant support for solid-phase peptide synthesis.

Modified support for solid-phase peptide synthesis which permits the synthesis of protected peptide fragments

Modified support for solid-phase peptide synthesis which permits the synthesis of protected peptide fragments