SPPS RESIN FOR HIGH PURITY PEPTIDE SYNTHESIS - Zalety

CEM Corporation

IDEAL ENVIRONMENT FOR PEPTIDE SYNTHESIS

The formation of secondary structures, either through intra- or inter-molecular aggregation, is a known problem in SPPS that is often encouraged by a hydrophobic resin environment1, such as a traditional polystyrene (PS) resin cross-linked with divinylbenzene (DVB). In contrast, SpheriTide® resin provides a more hydrophilic environment that contains amide bonds just like the growing peptide backbone, which discourages the peptide from aggregating with itself (intra-molecular aggregation). SpheriTide resin allows for higher purities and yields compared to non-polar resins where intra-molecular aggregation may limit the efficiency of the deprotection and/or coupling reaction.

BETTER PERFORMANCE AT HIGHER LOADING

The unique properties of the SpheriTide® resin allow for high purity syntheses of difficult peptides at extremely high loadings. Several peptides were prepared under identical synthesis conditions2 using a 1.04 mmol/g Rink Amide SpheriTide® (HL) resin and a 0.59 mmol/g Rink Amide MBHA polystyrene resin. As demonstrated by Glucagon 1-29 peptide3, the 1-27 fragment belonging to Z Domain of Protein A4, and PnIA(A10L) Conotoxin5 below, in each case, the SpheriTide (HL) resin outperformed the polystyrene resin.


IMPROVED SYNTHESIS PURITY WITH HL SPHERITIDE RESIN

Peptide UPLC Purity
0.59 mmol/g Rink amide MBHA 0.75 mmol/g Rink amide MBHA 1.04 mmol/g Rink amide SpheriTide
Influenza Virus Hemagglutinin
MEDSTYTKASKGC
57% 40% 55%
PnIA (A10L) Conotoxin
GCCSLPPCALNNPDYC
65% 51% 78%
1-27 Z Domain of Protein A
VDNKFNKEQQNAFYEILHLPNLNEEQR
59% 39% 73%
Glucagon
HSQGTFTSDYSKYLDSRRAQDFVQWLMNT
36% 18% 54%
† Standard HE-SPPS cycles (2,2,3 mL DMF washes) + 2 x 2min at 90 °C coupling for Arg. Glucagon used a 10 min 50 °C His coupling method. All samples lyophilized.



Glucagon 1-29 Peptide = HSQGTFTSDYSKYLDSRRAQDFVQWLMNT-NH2
Glucagon.jpgGlucagon with SpheriTide.jpg
 

GREATER REPRODUCIBILITY & HIGHER QUALITY PEPTIDES

Clustering of linker sites within a resin bead results in crowding and can lead to lower synthesis efficiency. Traditional polystyrene-based resins are prepared by heat-induced radical polymerization leading to random variations in cross-linking and functionalizing sites. In contrast, SpheriTide resins are highly structured. Cross-linking only occurs at the amine sites which are spaced with a minimum distance between sites, improving both peptide quality and batch-to-batch reproducibility between resin samples.
 

CONTROLLED SWELLING IN TFA

SpheriTide resins display controlled swelling in TFA. This can be an issue when using PEG-based resins which require large TFA/resin ratios for fully covering the resin.
 SpheriTide® is a registered trademark of Spheritech Ltd., is patented under PCT/EP2012/057264 and exclusively licensed by CEM Corporation..
[1] Fmoc Solid Phase Peptide Synthesis – A Practical Approach, W.C. Chan and P.D. White, Eds. 2000, Oxford University Press: New York. p. 30, 118. [2] All syntheses were performed using HE-SPPS methodology on the Liberty Blue peptide synthesizer (J.M. Collins, K.A. Porter, S.K. Singh, G.S. Vanier Org. Lett. 2014, 16, 940-943.) [3] J.S. Pedersen, D. Dikov, D.E. Otzen, Biochemistry 2006, 45, 14503-15512. [4] Simon et al. ChemBioChem 2014, 15, 713 - 720. [5] Abdel-Aal, A. M.; Papageorgiou, G; Quibell, M.; Offer, J. Chem Commun. 2014, 50, 8316 –8319.