The Peptide Synthesis/Antibody Production Core is one of the more recent additions to the DERC Support Facilities, funding being initiated on 3/1/92. This support service represents an expansion of a previously existing peptide synthesis unit which was created in 1986 by the Scientific Council at UMMC. Our goal has been to provide consultation and reliable, affordable services in the general areas of peptide synthesis, protein isolation and antibody production in an effort to enhance diabetes and endocrine work. Over the 9 months since its inception, this HPLC purifications, 55 amino acid analyses, 22 peptide conjugations and 47 antibody productions. By providing consultation and service in these areas at subsidized rates, we have greatly enhanced the potential for good research by each DERC investigator. Solid phase Merrified synthesis is performed using FAST-Moc chemistry on a scale of 0.2 mmol (200 mg for a 10 mer). This method permits cleavage from the resin-support by brief exposure to a mild acid, resulting in a cleaner product in a shorter time. The peptide is usually subjected to HPLC-purification and amino acid analysis is used for quantitation and to assay purity. Large-scale purifications of synthesized peptides (50- 200 mg) are performed using a 100 ml column of mu-Bondapak C-18 and a linear gradient in acetonitrile (2 hours/run). Pico-Tag analyses are performed by reverse-phase HPLC and pre-column derivatization using phenyl isothiocyanate. A synthetic peptides or other haptens are coupled to carrier molecules via NH2-, COOH, SH-or tyrosyl groups. These reactions can be designed so as to orient the hapten and direct antibody specificity.
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