Applications of microwave-assisted proteomics in biotechnology.
Biotechnology has recently celebrated 30 years both as a science and as a multi-billion dollar industry. One application of biotechnology is to use human genetic information to discover, develop, manufacture, and commercialize biotherapeutics. Recombinant proteins can be produced in large quantities at high purity. High-throughput proteomic analysis is at the heart of the biotechnology research and development process, and the industry is constantly striving to streamline and automate the analytical processes involved. Microwave-assisted proteomics has recently emerged as a tool for increasing the bio-catalysis of several processes including tryptic digestions lipase selectivities, identification of metal-catalyzed oxidation sites on proteins, identification of protein N- and C-termini and enzyme catalyzed N-linked deglycosylation. Here, we explore the above mentioned methods, and describe our experiences evaluating microwave-technology for other common proteomic protocols including: removal of N-terminal pyroglutamyl for antibody characterization, beta elimination and Michael addition for identification of phosphorylation sites on recombinant proteins and enzyme mediated O-linked deglycosylation.
Sandoval
Wendy N
WN
Protein Chemistry Department, Genentech Inc., South San Francisco, CA 94080, USA.
Pham
Victoria
V
Ingle
Elizabeth S
ES
Liu
Peter S
PS
Lill
Jennie R
JR
D000595:Amino Acid Sequence; D000906:Antibodies / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods* / Q000639:trends; D002384:Catalysis; D004798:Enzymes / Q000378:metabolism*; D006031:Glycosylation; D008049:Lipase / Q000378:metabolism; D008670:Metals / Q000737:chemistry; D008872:Microwaves; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010449:Peptide Mapping / Q000379:methods; D010766:Phosphorylation; D040901:Proteomics / Q000295:instrumentation / Q000379:methods* / Q000639:trends; D011751:Pyroglutamyl-Peptidase I / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014357:Trypsin / Q000378:metabolism
D000906:Antibodies; D004798:Enzymes; D008670:Metals; D008049:Lipase; D011751:Pyroglutamyl-Peptidase I; D014357:Trypsin