Construction Technician

BARBARA A. MCDONALD

Sudbury, MA ***** H: 978-***-**** / C: 314-***-**** *************@*******.***

Skilled Senior Research Associate with extensive academic and industry protein biochemistry experience.

Expert in a wide array of protein production, purification, and characterization techniques related to

recombinant proteins from bacterial cell culture and full-length proteins from sterile mammalian cell culture.

Further experience in molecular biology techniques including site-directed mutagenesis, DNA purification and

quantitation, and whole-genome library construction. Co-author of 11 publications in the field of Surfactant

Protein-D Biochemistry in the last eight years.

Areas of strength include:

Protein Production Protein/Antibody Purification Animal Handling

Experimental Troubleshooting Protein Characterization Team Collaboration

Sterile Tissue Culture Techniques Assay Development Data Analysis/Presentation

M ASTER OF S CIENCE – Biomedical Science: Northeastern University

B ACHELOR OF A RTS - Biochemistry: Canisius College

LABORATORY AND RESEARCH TECHNICIAN EXPERIENCE

WASHINGTON UNIVERSITY SCHOOL OF MEDICINE – Pathology Department – 2004 to 2012

Genome Sequencing Center – 2002 to 2004

Responsible for production, purification and extensive characterization of a large library of wild type

and mutant Surfactant Protein-D proteins from a variety of species. Produced in RosettaBlue

competent cells for recombinant proteins or sterile mammalian cell culture for full-length proteins.

Responsible for production, characterization and distribution of high quality, pure and stable variants

of Surfactant Protein-D to domestic and overseas research collaborators.

Evaluated Surfactant Protein-D wild type and mutant protein binding properties, affinity, oxidation

and interactions with ligands such as phosphatidylinositol, polysaccharides, and lipopolysaccharides.

Surface Plasmon Resonance used extensively to study the interaction of SP-D proteins with

components of influenza A virus.

Expert in the following Protein Biochemistry Techniques: AKTA FPLC Chromatography, His-Tag

Chelation Chromatography, Saccharide Affinity Chromatography, Gel-filtration Chromatography,

Biacore SPR and SensiQ SPR, SDS-PAGE, Western Blotting, Total Protein BCA Assays, Endotoxin Assays,

Dynamic Light Scattering, Spectrophotometry, and Silver Stain Assays.

Extensive experience in sterile tissue culture including protein production from mammalian cell lines,

cell line propagation and cloning, microscopy, screening, growth optimization, scale-up, harvest,

concentration and cryogenic cell storage.

Highly accomplished in ELISA binding assay and competition assay development.

Experience in DNA purification, DNA quantitation, and whole genome library construction

Skilled in Microsoft Word, Excel, PowerPoint, Sigma Plot (data analysis, graphing, and statistics),

Unicorn (for AKTA Purifier), Adobe PhotoShop, Adobe Illustrator, ImageQuant (image analysis of gels,

dot blots, and western blots), and Qdat (SensiQ SPR software).

UNIV. OF PITTSBURGH SCH OF MED - Dept. of Molecular Genetics and Biochemistry – 1990 to 1991

Dept. of Pathology – 1989 to 1990

Experienced in plasmid preparation techniques, restriction enzyme characterization, gel-

electrophoresis, chromium-release cytotoxicity assays, thymidine proliferation assays, gel-filtration

chromatography, SDS-PAGE, ELISA, spectrophotometry, animal handling, dissection, tissue culture.

CIBA-CORNING DIAGNOSTICS CORPORATION - Immunochemistry Department – 1983 to 1986

Product Development Laboratory – 1986-1989

Prepared, purified, and characterized protein-conjugate immunogens, acridinium-labeled antibodies,

and acridinium-labeled protein/nucleic acid antigens as well as immobilized antibodies to various

solid phases. Subsequently utilized these components to develop ELISA and chemiluminometric

screening assays for hybridoma selection. Antibodies commercialized in context of rapid, clinical

myocardial infarction testing.

Applied small animal and sterile tissue culture techniques to produce monoclonal antibodies including

immunizations,sera evaluation, dissection, cell fusions, cloning, microscopy, cryogenic cell storage,

and ascites production.

Extensive experience in antibody purification and characterization including HPLC, affinity

chromatography, ion-exchange chromatography, chemiluminometric assays, ELISA, RIA, SDS-PAGE,

total protein assays, spectrophotometry.

PUBLICATIONS

Crouch E, Nikolaidis N, McCormack F, McDonald B, Allen K, Rynkiewicz M, Cafarella T, White M, Lewnard K, Leymarie N, Zaia J, Seaton

B, Hartshorn K. Mutagenesis of SP-D informed by evolution and xray crystallography enhances defenses against Influenza A Virus in vivo.

J Biol Chem 2011:286:406**-*****.

Crouch EC, Hirche TO, Boxio R, Wartelle J, Benabid R, McDonald B, Heinecke J, Matalon S, Belaaouaj A. Myeloperoxidase-dependent

inactivation of Surfactant Protein D in vitro and in vivo. J Biol Chem 2010:285(22):167**-*****.

White MR, Boland P, Tecle T, Gantz D, Sorenson G, Tornoe I, Holmskov U, McDonald B, Crouch EC, Hartshorn KL. Enhancement of

antiviral activity of collectin trimers through cross-linking and mutagenesis of the carbohydrate recognition domain. J Innate Immun

2010:2(3):267-279.

Crouch E, Hartshorn K, Horlacher T, McDonald B, Smith K, Cafarella T, Seaton B, Seeberger PH, Head J, Recognition of mannosylated

ligands and influenza A virus by human SP-D: Contributions of an extended site and residue 343. Biochem 2009:48(15):3335-3345.

Matalon S, Shestra K, Kirk M, Waldheuser S, McDonald B, Smith, K, Gao Z, Belaaouaj A, Crouch EC, Modification of Surfactant Protein D

by reactive oxygen nitrogen intermediates is accompanied by loss of aggregating activity, in vitro and in vivo. FASEB J

2009:23(5):1415-1430 .

Cooley J, McDonald B, Accurso F, Crouch E, Remold-O’Donnell E, Patterns of neutrophil serine protease-dependent cleavage of

Surfactant Protein-D in inflammatory lung disease. J Leukoc Biol 2008:83(4):946-955.

Wang H, Head J, Kosma P, Brade H, Muller-Loennies S, Sheikh S, McDonald B, Smith K, Cafarella T, Seaton B, Crouch E, Recognition of

heptoses and the inner core of bacterial lipopolysaccharides by SP-D. Biochemistry 2008:47(2):710-720.

Crouch E, McDonald B, Smith K, Roberts M, Cafarella T, Seaton B, Head J, Critical role of Arg/Lys343 in the species-dependent

recognition of phosphatidylinositol by pulmonary Surfactant Protein D. Biochemistry 2007:46(17):5160-5169.

Crouch EC, McDonald B, Smith K, Cafarella T, Seaton B, Head J, Contributions of phenylalanine 335 to ligand recognition by human

Surfactant Protein D: Ring interactions with SP-D ligands. J Biol Chem 2006:281(26):180**-*****.

Crouch EC, Smith K, McDonald B, Briner D, Linders B, Holmskov U, Hartshorn K, Species differences in the carbohydrate binding

preferences of Surfactant Protein D: Contributions of aspartate-325 and multimeric assembly to ligand recognition Am J Resp Cell Mol

Biol 2006:35(1):84-94.

Crouch E, Tu Y, Briner D, McDonald B, Smith K, Holmskov U, Hartshorn K, Ligand specificity of human SP-D: Expression of a mutant

trimeric collectin that shows enhanced interactions with influenza A virus. J Biol Chem 2005:280(17):170**-*****.

Allen HJ, Karakousis C, Piver MS, Gamarra M, Nava H, Forsyth B, Matecki B, Jazayeri A, Sucato D, Kisailus E, Department of Surgical

Oncology, Roswell Park Memorial Institute, Buffalo, NY, Galactoside- Binding Lectin in Human Tissues. Tumor Biology 1987: 8(4):218-29.

Weetall HH, Forsyth B, Hertl W, A Direct Fuel Cell for the Production of Electricity from Ligand. Biotechnology and Bioengineering

1985: vol. 27.

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