UNC Biochemistry Core Facilities and Resources

 

 The biochemistry cores offer resources to investigators working on the molecular analysis of genes, proteins, peptides, cells and tissues.  Analytical approaches include mass spectrometry, NMR spectroscopy, x-ray crystallography and bioinformatics.

Immunology


CFAR Virology, Immunology & Microbiology Core 
provides support for research among CFAR (Center for AIDS Research) investigators and training in sample preparation and/or the performance of immunologic procedures.
Services include sample processing and storage, HLA-typing, CD4 counts, measurement of soluble biomarkers (ELISA, Luminex platform, ELISPOT), flow cytometry for phenotyping and functional analysis of immune cell populations; and antibody analysis.

 

Antibody Core Facility can help investigators produce monoclonal antibody-producing hybridoma cell lines. The core has partnered with an outside vendor (Precision Antibody) to work with investigator labs to develop hybridoma lines. Once developed, the core can take possession of these lines, store them for the investigators, and carry out large scale production and purification of the antibodies (multi-milligram). The facility also can produce large amounts of crude antibody in vitro for investigators (conditioned media) and purifies both polyclonal and monoclonal antibodies from supplied tissue culture media or ascites. If the Antibody Core is able to extract the antibody sequence from a hybridoma cell line, the Protein Expression and Purification Core  can produce it recombinantly using mammalian expression systems. This facility can also help produce protein antigen(s) for hybridoma projects.

 

Mass Spectrometry


The Biomarker Mass Spectrometry Facility provides expertise for qualitative and quantitative analysis of small molecular weight biomarkers including nucleic acids, peptides, and metabolites. In addition to LC/MS/MS analyses, they offer HPLC with UV detection and trace elemental analysis by ICP-MS.  Areas of focus for the facility have included DNA and protein adducts as markers of chemical exposure, oxidative stress, and endogenous DNA damage, characterization of chemical reaction products, identification and quantitation of metabolites, multi-element analysis and arsenic speciation. The services and analyses performed within the facility enable UNC investigators to study the molecular mechanisms of environmentally based diseases as well as the relationships between genetic and environmental factors.

 

 The Department of Chemistry Mass Spectrometry Core Laboratory specializes in small molecule mass spectrometry analysis.  The lab offers both independent operator and full-service mass spectrometry workflows.  The primary research focuses are organic and inorganic synthetic molecules, relative and absolute quantitation mass spectrometry, and trace metal analysis.
Research Applications include quantitative mass spectrometry, liquid chromatography separations, structural elucidation, MS/MS & MSn fragmentation mass spectrometry, complex mixture analysis, molecular formula confirmation, high resolution and accurate mass analysis, and trace metal determination/quantitation.

 
UNC Michael Hooker Proteomics Center provides researchers with state-of-the-art analysis of proteins from tissues, cells and other biological samples. The Center is also a resource for hands-on-training, education, and consultation in modern proteomics techniques. The Center offers an array of sample preparation and instrumentation services, as well as method development and in-depth consultation to help researchers optimize their experimental design.
Research applications include Identifying purified proteins and co-immunoprecipitated proteins, identifying proteins in from complex mixtures and quantify differences in protein abundance using labeled (SILAC, TMT) or label-free approaches. Identify of post-translational modifications such as acetylation, ubiquitylation, phosphorylation and sulfhydryl modificationsm, and Determining the exact mass of purified peptides and proteins.

 

Metabolomics


The Metabolomics Laboratory is a collaborative resource for global and targeted metabolite profiling from a wide range of biological samples.
The lab provides NMR analysis and absolute quantitation on nearly 300 metabolites including, amino acids, organic acids, amines, nucleic acids, carbohydrates, drug related metabolites, etc. from a variety of biological samples (such as urine, serum, CSF and saliva) cell and tissue extracts.Nuclear Magnetic Resonance (NMR)

 

Nuclear Magnetic Resonance (NMR)


The UNC Biomolecular NMR Laboratory supports research into the structure and dynamics of biological molecules.
Research applications include macromolecular structure and dynamics. Cryoprobes are available for small molecules and biological applications, including small molecule and natural product NMR.One-on-one training of new users is offered, as well as consultations to determine whether NMR will be useful in their research.

 

School of Pharmacy NMR provides a resource for small molecule and natural product NMR related to drug discovery.  Focus area: Small molecule and natural product NMR. Routine 1D and 2D 1H and 13C spectra, as well as NMR on 31P, 15N, and other nuclei. Macromolecular NMR experiments are available on the 500 MHz instrument.  Frequent training sessions are scheduled on demand. 

The Metabolomics Laboratory (by NMR) is a collaborative resource for global and targeted metabolite profiling from a wide range of biological samples.
The lab provides NMR analysis and absolute quantitation on nearly 300 metabolites including, amino acids, organic acids, amines, nucleic acids, carbohydrates, drug related metabolites, etc. from a variety of biological samples (such as urine, serum, CSF and saliva) cell and tissue extracts.Nuclear Magnetic Resonance (NMR)

 

Peptide Synthesis


High Throughput Peptide Synthesis and Array Facility
provides researchers with high quality consultation and peptide synthesis services including production, purification, and characterization of polypeptides and custom designed peptide arrays.
Research applications include synthetic peptides containing standard, post-translationally modified (e.g., acetylation, methylation, phosphorylation) and/or unnatural/nonstandard amino acids and fluorescent tags (e.g., 6-Cl-Trp, FAM-, TAMRA-) can be synthesized for a wide range of biological and biophysical applications, including FP, ITC, NMR, X-ray crystallography, mass spectrometry (with or without stable isotopes such as 2H, 13C, 15N), enzyme inhibition/competition assays, protein-protein interaction studies, cell permeable peptide reagents, antigen/antibody production including synthesis of Multiple Antigenic Peptides (MAPs), and high density peptide arrays.

 

Structural Biology


The Macromolecular Interactions Facility (Mac-In-Fac) provides instrumentation and resources for biophysical characterization of biological macromolecules and their interactions with cognate ligands.
Research applications include determination of solution molecular weight, oligomeric state and association/dissociation constants, sedimentation, diffusion and frictional coefficients, hydrodynamic and RMS radius, second virial coefficient, on  and off rates (kon and koff), stoichiometry, equilibrium binding constants and other thermodynamic parameters (∆G, ∆H and ∆S) of bimolecular interactions, secondary structure, melting profile and melting temperature (Tm).  
The Faclity provides one-on-one training of new users, consultations with prospective users to determine whether techniques available at the Mac-In-Fac will be useful in their research, and consultation in experimental design, data analysis and interpretation.


Macromolecular X-Ray Crystallography Core Facility
(MX) provides support and infrastructure necessary to enable individual investigators of any experience level to initiate and complete a macromolecular structural biology project.
Research applications include High-Throughput protein crystallization screening, monitoring, and crystal diffraction testing as well as data collection, crystal structure determination and analysis. Sample preparation may be done in the Protein Expression and Purification Core, which is located in the same facility.

 

The Protein Expression and Purification Core specializes in the production of pure, functional proteins for structural, biophysical, and biochemical studies. It is specifically designed as a “front-end” interface to other components of the Center for Structural Biology.  The services of this core are also applicable to other research areas, including antibody and antigen production and the manufacture of protein chips.

 

The R. L. Juliano Structural Bioinformatics Core Facility provides researchers with state-of-the-art consultation and collaboration in research studies requiring computational structural biology methods.
Bioinformatics analysis includes multiple sequence alignments, phylogenetic analysis, and ancestral sequence predictions. Protein structure analysis includes protein fold determination, structure 
superimposition and comparison, homology modeling, active site identification, mutation effects analysis, electrostatic surfaces, solvent accessibility calculations, subunit contact analysis, molecular dynamics calculations, free energies of binding, and docking of small molecules.

 

Protein Expression

The Protein Expression and Purification Core specializes in the production of pure, functional proteins for structural, biophysical, and biochemical studies. It is specifically designed as a “front-end” interface to other components of the Center for Structural Biology.  The services of this core are also applicable to other research areas, including antibody and antigen production and the manufacture of protein chips.

 

Analytical and Physical Chemistry

The Nanomedicines Characterization Core Facility (NCore) provides  comprehensive characterization of novel nanomedicines and nanomaterials.  Services include physicochemical assays (ICP-MS, Nanosight), comprehensive physicochemical characterization studies, and bioassays characterization services.
Physicochemical characterization (PC) of nanomaterials offers a comprehensive body of information that can be used to predict behavior of the nanomaterials in biological environments both in vitro and in vivo.  This information is essential in order to exercise tight control over inter-batch variations and maintain reproducibility of nanomaterials critical for their applications.

 

The CGIBD Advanced Analytics Core provides single cell genomic and proteomics analysis, high-throughput qPCR, and cost-effective immunoassays.
Single-cell analysis: single-cell mRNASeq, DNASeq, ATACSeq, specific target amplification (STA), and Western blotting. Both Fluidigm and DropSeq-based technologies are available for single-cell RNASeq, and can be used in tandem.
High-throughput qPCR: gene expression (GE) analysis, SNP genotyping, copy number variation, and quantitative digital PCR, with up to 9216 GE/SNP qRT-PCR or 36960 qdPCR values per run.
Cost-effective immunoassays: Luminex bead-based multiplex assays, ELISA, and DropArray “mini”-multiplexes, which use 80% less sample and reagent volume.