Microsequencing of Proteins

The ability to determine the amino acid sequence of a protein was a major advancement in understanding protein structure and function relationships. In the era of biotechnology, sequence analysis still occupies a position of primary importance. Common uses for protein sequence data include design of DNA probes and/or PCR primers, verification of the identity of expressed recombinant proteins, production of synthetic peptide antigens and characterization of post-translational protein modification. In addition, sequence data coupled with a computer search of protein databases has become very important in determining the novelty/identity of any isolated protein.

However, the major barrier to productive microsequencing projects is not the instrumentation but rather the successful purification of proteins/peptides suitable for analysis. Even the most sophisticated, sensitive instrument can not generate useful data from impure and/or inadequate amounts of protein. Unfortunately, most proteins of interest are available in very low abundance. Therefore, sample preparation is the most critical and challenging task in a successful microsequencing project today. The difficulties involved in the purification, storage and recovery of proteins are the major hindrances to the biochemical characterization and biological assay of low abundance proteins.

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