Molecular Biotechnology Core

Grant Information

The MBT Core offers consultation and services in the areas of customized peptide synthesis, peptide modification, purification and quality analysis, N-terminal protein sequencing by Edman degradation, CD Spectroscopy and biomolecular interaction analysis and equilibrium and kinetic measurements. These services are described in more detail below:

The peptide synthesis services include peptide design, synthesis, cyclization, HPLC purification and various types of modifications of synthetic peptides. Synthesis is carried out at 50 µmole, 100 µmole or 250 µmole scales on Omega 396 and ABI 431A synthesizers. The Core also synthesizes multiple antigen peptides (MAP). Quality is routinely confirmed by HPLC analysis and mass analysis by MALDI-TOF.

N-terminal Edman sequencing is performed on a Procise Model 492 Sequencer. To obtain an unambiguous sequence of an unknown protein about 5 pmol or more of protein / peptide is required. Purity and quantity are determining factors for successful sequencing. For proteins with blocked amino termini, the Core offers chemical or enzymatic digestion and separation of polypeptide fragments of such samples to obtain de novo N-terminal sequence information of the fragments.

The Core offers training and facilitates biomolecular interactions analysis using the Biacore 3000 system, based on the principle of Surface Plasmon Resonance (SPR). The Biacore 3000 is used for measuring the interactions of macromolecules with each other or with small ligands, and for equilibrium and kinetic measurements, competition assays, and epitope mapping. The core provides hands-on training on both Biacore 3000 and the BIAevaluation software to graduate students and fellows.

The Core maintains and provides training to use CD spectroscopy. CD Spectroscopy is a valuable instrument for rapid analysis of structural and conformational changes in a protein upon perturbation by mutation, temperature, pH and buffers. CD spectra arising due to peptide bond transitions in the "far-uv" spectral region (190-250 nm) provide information on secondary structure of protein, while CD spectra of proteins in the "near-uv" spectral region (250-350 nm) provide information on certain aspects of tertiary structure.