Reducing chemical background noise in automated protein sequencers.
Baseline noise resulting from the by-products of the automated Edman degradation of proteins is reduced using nonstandard coupling chemicals and dual-wavelength diode array detection of phenylthiohydantoin (PTH) amino acids. Changes to the chemistry include lowering the phenylisothiocyanate (PITC) concentration and using a novel coupling base diisopropylethylamine (DIPEA). The diode array is configured to cancel the UV absorbance of reaction by-products without adversely affecting the PTH signal intensities. Combining these techniques gives a significant reduction in chemical noise on analytical chromatograms. Less sample is required for sequence analysis since low-level amino acid peaks are more easily identified.
Farnsworth
V
V
Porton Instruments, Tarzana, CA 91356.
Carson
W
W
Krutzsch
H
H
D000595:Amino Acid Sequence; D002625:Chemistry, Organic / Q000295:instrumentation / Q000379:methods*; D007782:Lactoglobulins / Q000737:chemistry; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
D007782:Lactoglobulins; D011506:Proteins