Core A
PROTEIN BIOCHEMISTRY AND TISSUE ANALYSIS
Lawreen Heller Connors, PhD |
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Carl O’Hara, MD |
Core Leaders:
- Lawreen Heller Connors, Ph.D.
Assistant Professor (Research) of Biochemistry
- Carl J. O’Hara, M.D.
Professor of Pathology
The Protein Biochemistry and Tissue Analysis Core (Core A) has served as a resource providing centralized services to each of the three research projects in the original cycle of this program project study. Core A is a
- source for purified proteins,
- protein analysis and data reference center,
- tissue analysis laboratory and
- clinical specimens repository.
The purification of light chain (LC) proteins from clinical and recombinant sources is ongoing. Current separation protocols being used in Core Unit A to purify LCs are optimized versions of original column chromatographic and electrophoretic techniques. Automated equipment and updated technology using FPLC, HPLC and Phast PAGE instrumentation have dramatically increased efficiency and productivity. All purified proteins are assessed for homogeneity by gel electrophoresis, immunoblotting and HPLC. Other protein analysis services offered by Core Unit A include evaluation of specific LC size, hydrophobicity and the presence of glycosylated residues.
In addition, 2 dimensional gel electrophoresis analysis of serum, urine, fat and tissue samples is available in Core A. The compilation and distribution of sequence information (nucleotide and amino acid), protein structural data (primary, secondary, tertiary and quaternary) and experimental data is an important function of the core.
An important function of Core A is comprehensive tissue analysis. The evaluation of amyloid deposits in animal and human tissue is carried out using a variety of modalities including routine histological stains (H&E, PAS), special stains (Congo red, sulfated alcian blue), immunohistochemistry, electron microscopy and immunoelectron microscopy. The recent addition of a Leica polarizing light microscope and a SPOT digital imaging system attachment to Core Unit A enhances visualization and documentation of Congo red staining results.
The collection, cataloging and storage of clinical specimens including serum, bone marrow, urine, fat and other tissue (autopsy/biopsy) samples are ongoing. Repository records are continuously updated. Sample inventory information is being merged with relevant demographic data and clinical laboratory testing results (from Core C) for each case and organized in a computer database that is security-enforced to ensure confidentiality.
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