The University of British Columbia

RESEARCH INTERESTS

Purity and purification costs are becoming important issues in biotechnology as the industry matures and competitive products reach the marketplace. Research in my laboratory is focused on better understanding how biological macromolecules such as proteins and DNA interact with natural and unnatural ligands, and then developing new natural- and recombinant-protein purification processes based on high-affinity interactions between target proteins or drugs and separation media.

My laboratory runs the Centre for Biological Calorimetry, a federally funded centre which provides Canada with state-of-the-art methods for measuring the delicate energetics of biological interactions in solution or at interfaces. We have also been involved in the development of Raman spectroscopy, particularly fibre-optic probes and related instrumentation which allow remote acquisition of high-sensitivity UV-resonance Raman spectroscopy data for biological samples, including tissues, biological interfaces, and complex solutions.

In collaboration with Merck Chemicals (Germany), we have recently developed high resolution resins which utilize a layer of end-grafted polymeric chains to separate proteins on the basis of size and associated excluded-volume interactions. Our entropic-interaction chromatography columns, which are now commercially available, show dramatic separation performance for protein molecular weights ranging from a few kDa up to 1000 kDa.

When engineered properly, molecular genetics techniques provide a robust method for purifying recombinant proteins from the complex aqueous solutions in which they are produced. Our lab, in collaboration with Drs. Kilburn and Warren of the Microbiology department, is also interested in purification strategies which use the cellulose binding domains (CBD's) of Cellulomonas fimi cellulases as affinity tags. Genetic or chemical linkage of a CBD to the target protein creates a fusion protein which binds strongly to cellulose and retains the biological activity of the fusion partner. Recovery of the target protein is then achieved through either a modest change in system variables or enzymatic cleavage of the polypeptide backbone at the protein/CBD linkage.

Synthesis of pharmaceutical drugs often requires precursors of specified chirality. However, chemical syntheses of drug precursors, such as amino acids, typically yield racemic mixtures. We are involved in the development of large-scale, continuous processes for separating mixtures of chiral enantiomers. Research to date has involved the fabrication and characterization of ligands which discriminate between chiral enantiomers of therapeutic or industrial importance, and the development of systems for continuous separation of chiral enantiomers.

Research in my laboratories involves collaborations with industry, engineers, pathologists, chemists, and microbiologists. This collaborative research environment mirrors the multidisciplinary nature of industrial biotechnology and provides a sound foundation for understanding the complex structures and functions of proteins and thus, promising pathways for their purification.

RECENT SELECTED PUBLICATIONS

• Liu SM, Haynes CA, “Mesoscopic dynamic Monte Carlo simulations of the adsorption of proteinlike HP chains within laterally constricted spaces”, J Colloid Interface Sci. 282(2):283-92 (2005).
• Liu SM, Haynes CA, “Energy landscapes for adsorption of a protein-like HP chain as a function of native-state stability”,  J Colloid Interface Sci. 284(1):7-13 (2005).
• Lam H, Kavoosi M, Haynes CA, Wang DI, Blankschtein D. “Affinity-enhanced protein partitioning in decyl beta-D-glucopyranoside two-phase aqueous micellar systems”, Biotechnol Bioeng. 89(4):381-92 (2005).
• Pang P., Koska J, Coad B, Brooks D, Haynes CA, “Entropic interaction chromatography: Separating proteins on the basis of size using end-grafted polymer brushes”, Biotechnol Bioeng., [Epub ahead of print] Feb 10 (2005).
• Creagh AL, Tiong JW, Tian MM, Haynes CA, Jefferies WA, “Calorimetric studies of melanotransferrin (p97) and its interaction with iron”, J Biol Chem. [Epub ahead of print] Feb 9 (2005).
• Jervis E, Guarna MM, Doheny JG, Kilburn DG, and Haynes CA, “Dynamic Localization and Persistent Stimulation of Factor Dependent Cells by a Stem Cell Factor/Cellulose Binding Domain Fusion Protein”, Biotech. Bioeng., [Epub ahead of print], Mar 3 (2005).
• Lario PI, Pfuetzner RA, Frey EA, Creagh AL. Haynes CA, Maurelli AT, Strynadka CJ, “Structure and biochemical analysis of a secretin pilot protein”, EMBO Journal, in press, Feb. 23 (2005).
• Kavoosi, M, Meijer J, Kwan E, Creagh AL, Kilburn DG, Haynes CA, “Inexpensive one-step purification of polypeptides expressed in E. coli as fusions with the family 9 carbohydrate-binding module of xylanase 10A from T. maritima”, J. Chromatography B, 807: 82-89 (2004).
• So AP, Turner RFB, Haynes CA, “Increasing the efficiency of SAGE adaptor ligation by directed ligation chemistry”, Nucleic Acids Research (Methods Online), 32(12):e96 (2004).
• Liu SM, Haynes CA, “Mesoscopic analysis of conformational and entropic contributions to nonspecific adsorption of HP copolymer chains using dynamic Monte Carlo simulations”, J Colloid Interface Sci. 275(2):458-69 (2004).
• Rodriguez B, Kavoosi M, Jürgen Koska J, Creagh AL, Kilburn DG, Haynes CA, “Inexpensive and generic affinity purification of recombinant proteins using a family 2a CBM fusion tag”, Biotechnology Progress, 20(5):1479-89 (2004).
• Bekker EG, Creagh AL, Sanaie N, Yumoto F, Lau GHY, Tanokura M, Haynes CA, Murphy MEP, “Specificity and function of the synergistic anion in ferric binding protein from Neisseria gonorrhoeae: roles of Gln58 and Gly140 in iron binding”, Biochemistry, 43(28):9195-203 (2004).
• Haynes C, “The importance of bioseparations: giving credit where credit is due”, Biotechnol Bioeng. 87(3):257-8 (2004).
• Tsai M, Sampaleanu LM, Greene C, Creagh AL, Haynes CA, Howell PL, “A duck d1 crystallin loop mutant provides insight into residues important for argininosuccinate lyase activity”, Biochemistry, 43: 11672-682 (2004).
• Haynes CA, “Biopartitioning and Purification”, J Chromatogr B Analyt Technol Biomed Life Sci. 807(1):1 (2004).
• Haynes CA, “ Highlights of the 12th International Conference on Biopartitioning and Purification”, J. Chromatography B, special issue Proceedings of the 12th International Conference on Biopartitioning and Purification, (CA Haynes, guest editor) 807: 5 (2004).
• Haynes CA, “The importance of bioseparations: giving credit where credit is due”, Biotechnology & Bioengineering, special issue Advances in Bioseparations (CA Haynes, editor) 86(9): 1117-1118 (2004).
• J. Koska and C.A. Haynes, "Performance and Modelling of a Ligand-Exchange-Based Liquid-Liquid Extraction System for Chiral Separations", AIChE J., 49, 82-89 (2003).
• Jianhua Hu, Amy H.Y. Wu, Candy M.W. Chuang, Mark G.M. Chen, Eric G.M. Yee, Takehiko Ichioka, Keiko Nishikawa, Charles Haynes, and Yoshikata Koga, "Chemical Potential and Concentration Fluctuation in some aqueous alkane-mono-ols at 25 C", Canadian Journal of Chemistry, 81(2), 141-149 (2003).
• C.A. Haynes, J.-K. Hui, S. Lambert, "A System for Multiplex Sequencing and Hybridization-Based Pullout of DNA Extension Products", Nucleic Acids Research (Methods Online), 31: e41 (2003).

OTHER AFFILIATIONS/MEMBERSHIPS

• Associate Professor - Biotechnology Laboratory

• NATO Postdoctoral Fellow, 1992-93

   

TEACHING

• CHBE 453 - Biotechnology Process Design Project

• CHBE 481 - Advanced Topics in Biological Engineering

• CHBE 533 - Bioanalytical Chemistry

• APSC 122 - Introduction to Engineering