Comparative evaluation of four trityl‐type amidomethyl polystyrene resins in Fmoc solid phase peptide synthesis

Zikos, Christos; Livaniou, Evangelia; Leondiadis, Leondios; Ferderigos, Nikolas; Ithakissios, Dionyssis S.; Evangelatos, Gregory P.

doi:10.1002/psc.454

Cited by 17 publications

(9 citation statements)

References 28 publications

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“…All laboratory solvents and chemicals were purchased from Merck (Darmstadt, Germany) or Sigma (St. Louis, MO) and they were analytical grade. , and ProTa [101][102][103][104][105][106][107][108][109] were synthesized by the Fmoc-solid phase peptide synthesis following a protocol that has been previously described (Zikos et al 2003). The peptides synthesized were purified on a Waters 600E HPLC System (Waters; Milford, MA) by semipreparative reverse phase-high performance liquid chromatography (RP-HPLC) using a 250 3 12.7-mm (ID) Nucleosil 7 C18 column (Macherey Nagel; Dueren, Germany).…”

Section: Methodsmentioning

confidence: 99%

Immunocytological and Preliminary Immunohistochemical Studies of Prothymosin α, a Human Cancer–associated Polypeptide, With a Well-characterized Polyclonal Antibody

Klimentzou

Drougou

Fehrenbacher

et al. 2008

J Histochem Cytochem.

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Prothymosin α (ProTα) is a nuclear polypeptide of great biological and, possibly clinical, importance, because its expression levels have been associated with early diagnosis/prognosis of human cancer. It is therefore interesting to raise easily available and cost-effective antibodies that would be applied to develop reliable ProTα immunodiagnostics. In this study, New Zealand white rabbits and laying hens were parallel immunized against intact ProTα or the synthetic fragments ProTα[1-28], ProTα[87-109], and ProTα[101-109], all conjugated to keyhole limpet hemocyanin (KLH). The corresponding antibodies G and Y were immunochemically evaluated in parallel with ELISA and Western blot systems and applied to fluorescence immunocytology experiments using various cancer cell lines and normal cells. The antibody G raised against ProTα[101-109]/KLH had excellent functional characteristics in the Western blot and immunocytology experiments, where the fluorescent signal was almost exclusively shown in the cell nucleus independently of the cells assayed. The above antibody has been applied to preliminary IHC staining of human cancer prostate tissues, leading to a high percentage of clearly and intensively stained nuclei in the adenocarcinoma tissue; this antibody can be further used in cancer tissue immunostaining and in research concerning the role of ProTa in tumorigenesis.

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Section: Methodsmentioning

confidence: 99%

Immunocytological and Preliminary Immunohistochemical Studies of Prothymosin α, a Human Cancer–associated Polypeptide, With a Well-characterized Polyclonal Antibody

Klimentzou

Drougou

Fehrenbacher

et al. 2008

J Histochem Cytochem.

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“…Synthesis of ADNF‐9 peptide and its functionalized derivatives was performed according to solid‐phase strategy (16). Both peptides (Figure 1) were labeled with FITC for tracking their intracellular uptake.…”

Section: Resultsmentioning

confidence: 99%

“…ADNF‐9 peptide derivatives were synthesized on an in‐house‐prepared polystyrene resin (0.5 mmol/g) (Figure 1) (16). Solid‐phase peptide synthesis was performed manually.…”

Section: Methodsmentioning

confidence: 99%

Cholesteryl‐Functionalized ADNF‐9 Peptide: Enhanced Membrane Transport Through Mouse Neuroblastoma Neuro‐2a Cells

et al. 2012

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A cholesteryl‐functionalized derivative of activity dependent neurotrophic factor‐9 peptide (a nine amino acid core peptide of activity‐dependent neurotrophic factor, acting against Alzheimer’s disease) was synthesized aiming at the improvement of its bioavailability. Therefore, its uptake was comparatively investigated with that of its parent peptide by employing mouse neuroblastoma Neuro‐2a cells. Owing to the hydrophobic character of this cholesteryl‐functionalized peptide, it exhibited enhanced permeability and intracellular uptake while it also retained its low cytotoxicity at concentrations up to 1 μm. FACS analysis also revealed that when Neuro‐2a cells were treated with this activity dependent neurotrophic factor‐9 derivative, at a concentration of 50 nm, an almost 100% uptake was obtained. In addition, in vitro biological activity experiments showed that the functionalized peptide retained its neurotrophic activity at femtomolar concentration range.

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“…The Fmoc-protected aminocaproic acid (Fmoc-Aca-OH) was prepared in-house. The 4-CN-tritylamidomethyl polystyrene resin used in the synthesis of the "lysine dendrimer" was prepared in-house, as previously described [12]. DMF, DCM, THF, TFA, HPLC solvents (analytical grade), and other common laboratory chemicals were obtained from Merck or Sigma.…”

Section: Methodsmentioning

confidence: 99%

Solid-Phase Synthesis of a Biotin Derivative and its Application to the Development of Anti-Biotin Antibodies

Papasarantos

Klimentzou

Koutrafouri

et al. 2009

Appl Biochem Biotechnol

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A biotin derivative, namely biotin-aminocaproic acid-lysine (BAL), was synthesized with solid-phase chemistry, conjugated to a carrier-protein, and used for rabbit immunization. The aminocaproic acid-lysine "long-arm" was used in order to project the biotin-hapten above the carrier-protein surface. Lysine was selected due to its N(epsilon)-amino group, through which BAL was conjugated to the carrier-protein. BAL was synthesized on a commercially available resin with the Fmoc-solid-phase strategy; this has simplified the experimental procedure, overcome the need for intermediate purification steps, and led to a final product of high purity, with high yield. The anti-BAL antibodies recognized free biotin, as shown with an in-house-developed ELISA, in which biotin conjugated to a synthetic "lysine-dendrimer" was used to coat the ELISA microwells. In immunocytology and Western-blot experiments, the anti-BAL antibodies led to similar results with those obtained with streptavidin. Synthetic derivatives of hapten molecules that can be easily prepared with solid-phase chemistry, such as BAL, may be used for the development of specific antibodies for the corresponding hapten.

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Comparative evaluation of four trityl‐type amidomethyl polystyrene resins in Fmoc solid phase peptide synthesis

Cited by 17 publications

References 28 publications

Immunocytological and Preliminary Immunohistochemical Studies of Prothymosin α, a Human Cancer–associated Polypeptide, With a Well-characterized Polyclonal Antibody

Immunocytological and Preliminary Immunohistochemical Studies of Prothymosin α, a Human Cancer–associated Polypeptide, With a Well-characterized Polyclonal Antibody

Cholesteryl‐Functionalized ADNF‐9 Peptide: Enhanced Membrane Transport Through Mouse Neuroblastoma Neuro‐2a Cells

Solid-Phase Synthesis of a Biotin Derivative and its Application to the Development of Anti-Biotin Antibodies

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