Biomedical Analysis Lab

The Biomedical Analysis Lab (BAL) is a core lab involved in a variety of both basic and clinical research projects. We provide services to investigators at multiple Nemours Children's sites, as well as to external collaborators, including:

• Support and consultation on experimental design
• Sample analysis
• Data interpretation
• Training and mentoring of junior faculty, students and fellows
• Generation of preliminary data for investigators’ grant applications

The lab applies a multidisciplinary approach to the exploration of various changes in the human body at the molecular and cellular levels. To follow these changes in children, the lab uses state-of-the-art techniques, including:

• Stable isotope (nonradioactive) dilution techniques
• Mass spectrometry (a unique and powerful technique that is available at only a few centers in the United States)
• Assays based on high-performance liquid chromatography, electrophoresis and multiplex assay systems

One area of focus is obesity-related biomarkers for cardiometabolic disease (CMD). The lab uses several contemporary techniques that are especially powerful in studying biomarkers and following the changes in health and disease conditions in children. The studies the lab performs will lead to better understanding of the disease conditions and the development of more directed therapies. We collaborate with investigators from Oregon to Brazil, including the Mayo Clinic College of Medicine and ChristianaCare. The lab presented data at national and international meetings.

Current Research Group

• Prabhakaran (Babu) Balagopal, PhD, Director
• Shawn Sweeten, BS, Laboratory Manager
• Astride Altomare, BS, Senior Research Assistant
• Karl J. Mann, BS, Senior Research Assistant

We provide services to investigators at multiple Nemours sites, as well as to external collaborators, including:

Services

• Mass spectrometry (proteins, amino acids, and metabolites), including GCMS, IRMS and liquid chromatography tandem mass spectrometry (LC-MS/MS)
• HPLC assays for various peptides and metabolites
• Enzyme-linked immunosorbent assay (ELISA)
• Radio-immunoassays (RIA)
• Electrophoresis
• Nephalometry
• Immune-affinity chromatography
• Multiplex assay systems

Europa Hydra 20/20 Gas Chromatograph - Pyrolysis - Isotope Ratio Mass Spectrometer (GC-P-IRMS)

Europa 20-20 isotope ratio mass spectrometer, interfaced with Agilent/6890 GC with HP7683 autosampler and attached electromagnet analyzer; mass range is m/z = 1-70 at 3 kV. This has both pyrolysis and combustion capability (GC-C/P-IRMS) and a module for breath analysis. Two different pyrolysis interfaces between the GC and IRMS provide sensitive measurement of 2H and 18O. Triple Faraday collector system for measuring the isotopes of 13C, 18O, or 15N from CO2, CO, or N2 gases.

Nephelometer (BN II Nephelometer, Dade-Behring)

This instrument is used mainly for the high-sensitivity measurement of concentrations of an array of plasma proteins such as CRP, fibrinogen, RBP4, transthyretin, Apo A1, ApoB-100 and complement proteins.

Micromass Optima Isotope Ratio Mass Spectrometer

Interfaced with a Hewlett Packard 5890 GC with combustion capability (GC-C-IRMS) and equipped with a Gilson auto-sampler for breath analysis.

Hewlett Packard HP-5973 Gas Chromatograph — Mass Spectrometer (GCMS)

HP6890 GC with an HP7683 autosampler; Quadruple mass analyzer; m/z = 0-800 mass range; electron impact (EI) ionization; nNegative (NCI) and positive chemical ionization (PCI). Used for measuring stable isotopic enrichments in metabolites of biological samples in the range of 0.5 -10 mole % excess (mpe) tracer by selected ion monitoring (SIM).

Bio-Rad Prep Cell system

For the purification of specific muscle proteins such as (myosin heavy chain).

Immununo-affinity chromatography system (Bio-Rad)

For the purification of various plasma proteins.

Gilson HPLC

For the identification and separation of metabolites, peptides and proteins in biological samples. This HPLC is equipped with an ESA electrolytic conductivity detector (ECD), a Gilson UV-Vis detector and a Gilson Fluorometer.

Spectraphysics HPLC

For the identification and separation of metabolites, peptides and proteins in biological samples. This HPLC system has UV-Vis detection with an SP 8880 autosampler and a Pharmacia LKB Frac-200 fraction collector.