Department of Chemical & Biomolecular Engineering
College of Engineering, University of Nebraska–Lincoln
207N OTHM Lincoln NE 68588-0643
Current research interests may be described using the recently coined phrase "Functional Proteomics," but with a twist. In this case, the object is not looking at thousands of different proteins that cells are producing. Instead, our research is looking at recombinant proteins that are produced in bioreactors. These proteins are very complex because they have numerous post-translational modifications (PTMs); i.e., chemical modifications made to the amino acid side chains, such as glycosylation, gamma-carboxylation, phosphorylation, sulfation, and proteolytic processing. Thus, in the lab a "pure" protein can have many different subpopulations that differ in the nature and/or extent of their modification. The objective of this research is to characterize the structure of these subpopulations and then relate the structure to the in vivo function and pharmacokinetic properties. This work requires expertise in protein purification and analytical biochemistry, which is carried out in a Protein Characterization Laboratory (PCL), part of the Biological Process Development Facility at UNL. HPLC, SDS PAGE methods, enzymatic assays, peptide mapping, and LC-MS-MS are routinely used as tools for this work. Presently being used is an Applied Biosystems 4000 QTrap MS instrument for these molecules. This MS instrument has been invaluable in helping us discern the PTMs on the proteins.
N-glycosylation microheterogeneity and site occupancy of an Asn-X-Cys sequon in plasma-derived and recombinant protein C, Geun-Cheol Gil, William H. Velander, and Kevin E. Van Cott
Haemophilic Factors Produced by Transgenic Livestock: Abundance that can enable alternative therapies worldwide, Kevin E. Van Cott, Paul E. Monahan, Timothy C. Nichols, and William H. Velander
Inside-out crosslinked and commercial-scale hydrogels, and sub-macromolecular selective purification using the hyrdogels, William H. Velander and Kevin E. Van Cott
Affinity Purification of Biologically Active andInactive Forms of Recombinant Human Protein C Produced in Porcine Mammary Gland, Kevin E. Van Cott; Barry Williams Department of Chemical Engineering,Virginia Tech University, Blacksburg, VA 24061, USA; William H. Velander; Francis C. Gwazdauskas; Tim Lee Jerome Holland Laboratory, American Red Cross, Rockville, MD 20855, USA; Henryk Lubon; and William N. Drohan