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  • Design, synthesis and evaluation of a gonadotropin releasing hormone-based subunit vaccine in rams (Ovis aries).

Design, synthesis and evaluation of a gonadotropin releasing hormone-based subunit vaccine in rams (Ovis aries).

Vaccine (2015-02-14)
Daryn Goodwin, Pegah Varamini, Pavla Simerska, Michael J D'Occhio, Istvan Toth
ABSTRACT

Immunocastration using gonadotropin-releasing hormone (GnRH)-based vaccines has been investigated in rams to reduce aggressive and sexual behaviour and to control meat quality. Despite considerable efforts, a practical GnRH vaccine has yet to be developed for rams. In the present study, a A GnRH-lipopeptide vaccine (GnRH-LP) including two copies of GnRH, 2-amino-d,l-hexadecanoic acid (C16), and a unique T helper epitope, was examined in rams. Rams received a primary and secondary vaccination of GnRH-LP without additional adjuvant (Group 1) or with the adjuvant AdjuVac™ (Group 2). In both Group 1 and 2 anti-GnRH antibody titres increased after secondary vaccination, however, the antibody titres were higher (p<0.01) for rams in Group 2. The latter rams showed a marked decrease in testicular size. The marked and sustained reduction in testicular size in rams treated with GnRH-LP+AdjuVac™ provides the basis for an effective immunocastration vaccine in rams.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Water-16O, ≥99.94 atom % 16O
Sigma-Aldrich
Dichloromethane, anhydrous, ≥99.8%, contains 40-150 ppm amylene as stabilizer
Sigma-Aldrich
Acetonitrile, anhydrous, 99.8%
Sigma-Aldrich
N,N-Diisopropylethylamine, purified by redistillation, 99.5%
Supelco
Acetonitrile, analytical standard
Supelco
Dichloromethane, Selectophore, ≥99.5%
Supelco
Dichloromethane, analytical standard
Supelco
Trifluoroacetic acid, analytical standard
Sigma-Aldrich
N,N-Diisopropylethylamine, ReagentPlus®, ≥99%
Sigma-Aldrich
Benzhydrylamine, 97%
Sigma-Aldrich
Dichloromethane, suitable for HPLC, ≥99.9%, contains 40-150 ppm amylene as stabilizer
Sigma-Aldrich
Acetonitrile, electronic grade, 99.999% trace metals basis
Sigma-Aldrich
Acetonitrile, ≥99.5% (GC)
Sigma-Aldrich
Trifluoroacetic acid, ReagentPlus®, 99%
Supelco
Methylene Chloride, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Acetonitrile, ReagentPlus®, 99%
Sigma-Aldrich
Trifluoroacetic acid, puriss. p.a., suitable for HPLC, ≥99.0% (GC)
Sigma-Aldrich
Dichloromethane, contains 40-150 ppm amylene as stabilizer, ACS reagent, ≥99.5%
USP
Residual Solvent Class 2 - Acetonitrile, United States Pharmacopeia (USP) Reference Standard
Methylene chloride, European Pharmacopoeia (EP) Reference Standard
USP
Methyl alcohol, United States Pharmacopeia (USP) Reference Standard
USP
Glacial acetic acid, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Acetonitrile, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.5% (GC)
Sigma-Aldrich
Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)
Sigma-Aldrich
Dichloromethane, puriss., meets analytical specification of Ph. Eur., NF, ≥99% (GC)
Sigma-Aldrich
N,N-Diisopropylethylamine, 99.5%, biotech. grade
Sigma-Aldrich
Acetonitrile, biotech. grade, ≥99.93%
Sigma-Aldrich
Acetonitrile, ACS reagent, ≥99.5%
Sigma-Aldrich
Dichloromethane, biotech. grade, 99.9%, contains 40-150 ppm amylene as stabilizer
Sigma-Aldrich
Dichloromethane, ACS reagent, ≥99.5%, contains 40-150 ppm amylene as stabilizer