Peptide synthesis. Part 2. Procedures for solid-phase synthesis using N<sup>α</sup>-fluorenylmethoxycarbonylamino-acids on polyamide supports. Synthesis of substance P and of acyl carrier protein 65–74 decapeptide

Atherton, Eric; Logan, C.; Sheppard, R. C.

doi:10.1039/p19810000538

Cited by 416 publications

(187 citation statements)

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“…Thirty-five 15-mer peptides from hepatitis C virus core protein genotype 1b [35] were synthesized manually by the solid phase method of Merrifield [36] using the Fmoc alternative [37] and a multiple solid phase peptide synthesizer [38]. Peptide HA(306 -320) (CPKYVKQNTL KLATG) from Influenza A/Texas/77 virus hemagglutinin was synthesized manually, and biotinylated (while still attached to the resin) with an excess of N-hydroxysuccinimide-conjugated biotin (NH-LC-biotin) (Pierce).…”

Section: Materials and Methods Peptide Synthesismentioning

confidence: 99%

Specific and general HLA-DR binding motifs: comparison of algorithms

et al. 2000

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Using panels of peptides well characterized for their ability to bind to HLA DR1, DRB1*1101, or DRB1*0401 molecules, algorithms were deduced to predict binding to these molecules. These algorithms consist of blocks of 8 amino acids containing an amino acid anchor (Tyr, Phe, Trp, Leu, Ile, or Val) at position i and different amino acid combinations at positions i؉2 to i؉7 depending on the class II molecule. The sensitivity (% of correctly predicted binder peptides) and specificity (% of correctly predicted non-binder peptides) of these algorithms, were tested against different independent panels of peptides and compared to other algorithms reported in the literature. Similarly, using a panel of 232 peptides able to bind to one or more HLA molecules as well as 43 non-binder peptides, we deduced a general motif for the prediction of binding to HLA-DR molecules. The sensitivity and specificity of this general motif was dependent on the threshold score used for the predictions. For a score of 0.1, the sensitivity and specificity were 84.7% and 69.8%, respectively. This motif was validated against several panels of binder and non-binder peptides reported in the literature, as well as against 35, 15-mer peptides from hepatitis C virus core protein, that were synthesized and tested in a binding assay against a panel of 19 HLA-DR molecules. The sensitivities and specificities against these panels of peptides were similar to those attained against the panels used to deduce the algorithm. These results show that comparison of binder and non-binder peptides, as well as correcting for the relative abundance of amino acids in proteins, is a useful approach to deduce performing algorithms to predict binding to HLA molecules.

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Section: Materials and Methods Peptide Synthesismentioning

confidence: 99%

Specific and general HLA-DR binding motifs: comparison of algorithms

et al. 2000

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“…Affinity purified anti-rabbit immunoglobulin G conjugated to alkaline phosphatase was from Promega (Sydney, NSW, Australia). Synthetic Cterminally amidated peptides were synthesised by the Fmoc procedure [9]. The first peptide, designated Pb, was homologous to the fl-secretase cleavage site in APP695 between residues 588 and 601 (TEE-ISEVKMDAEFR-NHz).…”

Section: Methodsmentioning

confidence: 99%

Gelatinase A possesses a β‐secretase‐like activity in cleaving the amyloid protein precursor of Alzheimer's disease

et al. 1995

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“…Peptides were synthesized by the solid phase method of Merrifield (21) using the Fmoc alternative (22). The synthesis was done manually and the ninhydrin test of Kaiser (23) was used to monitor every step.…”

Section: Peptide Synthesismentioning

confidence: 99%

Induction of cytotoxic T lymphocytes in mice against the principal neutralizing domain of HIV-1 by immunization with an engineered T-cytotoxic-T-helper synthetic peptide construct

Lasarte

Sarobe

Gullón

et al. 1992

Cellular Immunology

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Peptide constructs were engineered by colinear synthesis of two short synthetic peptide determinants: a determinant recognized by T helper cells (TD,) and a determinant recognized by T cytotoxic cells (TD,). Three types of constructs were synthesized: TD,-TDr, , TD,-TD, , and TD,,-KK-TD,, where KK are two lysine residues. In vivo immunization with free construct induced cytolytic lymphocytes (CTL) only in the case of TD,-TD,,. However, immunization with spleen cells to which these constructs had been internalized by hypertonic shock, induced CTL activity in all three cases. No CTL could be induced after immunization with free TD, in either protocol. These results indicate that cell internalization of the construct might be essential for CTL induction, and also, that "help" from the TDr, seems to be required. o 1992 Academic PKSS, IX.

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Peptide synthesis. Part 2. Procedures for solid-phase synthesis using N^α-fluorenylmethoxycarbonylamino-acids on polyamide supports. Synthesis of substance P and of acyl carrier protein 65–74 decapeptide

Cited by 416 publications

References 1 publication

Specific and general HLA-DR binding motifs: comparison of algorithms

Specific and general HLA-DR binding motifs: comparison of algorithms

Gelatinase A possesses a β‐secretase‐like activity in cleaving the amyloid protein precursor of Alzheimer's disease

Induction of cytotoxic T lymphocytes in mice against the principal neutralizing domain of HIV-1 by immunization with an engineered T-cytotoxic-T-helper synthetic peptide construct

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Peptide synthesis. Part 2. Procedures for solid-phase synthesis using Nα-fluorenylmethoxycarbonylamino-acids on polyamide supports. Synthesis of substance P and of acyl carrier protein 65–74 decapeptide

Cited by 416 publications

References 1 publication

Specific and general HLA-DR binding motifs: comparison of algorithms

Specific and general HLA-DR binding motifs: comparison of algorithms

Gelatinase A possesses a β‐secretase‐like activity in cleaving the amyloid protein precursor of Alzheimer's disease

Induction of cytotoxic T lymphocytes in mice against the principal neutralizing domain of HIV-1 by immunization with an engineered T-cytotoxic-T-helper synthetic peptide construct

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Product

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Peptide synthesis. Part 2. Procedures for solid-phase synthesis using N^α-fluorenylmethoxycarbonylamino-acids on polyamide supports. Synthesis of substance P and of acyl carrier protein 65–74 decapeptide