Multipin peptide synthesis at the micromole scale using 2‐hydroxyethyl methacryiate grafted polyethylene supports

Abstract

The multipin peptide synthesis procedure has been adapted to allow the synthesis of peptides at micromole loadings. The original solid pin support was replaced with a detachable crown-shaped polyethylene support with an increased surface area. In addition, the polyethylene crowns were radiation-grafted with 2-hydroxyethyl methacrylate monomer instead of acrylic acid to yield hydroxy functionalised supports with a larger concentration of polymer and hence a larger peptide capacity. Fmoc-/3-Alanine was directly esterified to the HEMA hydroxy groups with subsequent addition of a diketopiperazine-forming handle for peptide attachment. Peptides varying in length from 10 to 25 residues were assembled at a number of loadings from 1.0 to 2.2 p o l . Purity of peptides at all loadings was equal to, and in some instances superior to, that achieved on conventional solid-phase supports. 0 Munksgaard 1993.

High Throughput T Epitope Mapping and Vaccine Development

“…In fact, peptides are currently used at 0.1–10 ug/mL in culture. Miniaturized peptide synthesis on pins [71–73] or on paper spots [74, 75] permits a less expensive peptide synthesis. In addition, microsynthesis in 96 well plate formats permits an easier transfer of peptides to 96 well or to 384 well plates, thereby abating the risk of positional errors or cross-contaminations.…”

Combinatorial/Library Peptide Synthesis

“…Simply dipping these pins into the wells with appropriate activated building blocks (amino acids) allows for simultaneous parallel synthesis. The material, shape, and functionalization of the pins went through multiple generations [66][67][68][69][70][71][72][73][74][75][76][77][78][79][80][81]. The optimized version of the multipin technology uses SynPhaseÔ lanterns or crowns -polymeric barrels in a shape of miniature Chinese lanterns or crowns [82,83] (Figure 12.1).…”

Efeito nociceptivo induzido por fosfolipases A2 (FLA2 variantes Lys49 e Asp49) isoladas do veneno de serpentes Bothrops asper: caracterização dos mecanismos centrais e determinantes moleculares

“…Estudo de neutralização de sítios catiônicos pela heparina. Baseados em dados sugerindo que a região catiônica 115-129 da miotoxina II (FLA 2 variante Lys49) é a responsável pela sua atividade citolítica e miotóxica (Lomonte et al, 1994b;Nunez et al, 2001), foram utilizados, para a avaliação da atividade hipernociceptiva, peptídeos 13-mer, sintetizados pela estratégia Fmoc [N-(9-fluorenil) metoxicarbonil] (Valerio et al, 1993). Foram empregados: a) o peptídeo p115-129 (sistema de numeração (Renetseder et al, 1985) correspondente aos resíduos 105-117 (KKYRYYLKPLCKK) da miotoxina II (Francis et al, 1991) Guo et al, 1995;Andoh et al, 1998;Okano et al, 1998).…”