Growth hormone, tryptic digest

Hancock, W.S., Chloupek, R.C., Kirkland, J.J., Snyder, L.R. (1994). Temperature as a variable in reversed-phase high-performance liquid chromatographic separations of peptide and protein samples. I. Optimizing the separation of a growth hormone tryptic digest. J. Chromatogr. A 686, 31 -3. 

Recombinant human growth hormone, tryptic digests Vydac 218TPB5 (Cl8), 5 pm, 300 A A 0.1% TFA/water, B 0.09% TFA/acetonitrile, gradient from 0 to 60% B 250 mm x 100 pm i.d. Electrically assisted capillary HPLC, coupling with electrospray ionization-mass spectrometry... 

Peptide mapping (the separation of the tryptic digest) of /i-lactoglobulin and human growth hormone (hGH). 

Frenz et al. applied the concept of trace enrichment to the tryptic digest of recombinant human growth hormone (rhGH). One of the innovations they introduced [39] was to directly monitor the... 

R. C. Chloupek, W. S. Hancock, and L. R. Synder, Computer simulation as a tool for the rapid optimization of the HPLC separation of a tryptic digest of human growth hormone, J. Chromatogr., 594 65 (1992). 

A recent modification of the atmospheric pressure ionization technique involving a special low dead volume interface was described by Thomson etal. [27]. They employed packed microbore columns (170 p, 320 p, and 500 p I. D. with lengths ranging from 5 to 15 cm) in conjunction with a low-volume, wall-coated capillary column as an interface. The total ion current chromatogram of the tryptic digest sample of about 1 picomole of human growth hormone is shown in figure 29. The column was packed with an octadecyl bonded phase... 

Spectmm of a Product from the Tryptic Digest of Human Growth Hormone Obtained from a Low Dead Volume Atmospheric Ionization Interface Courtesy of the Perkin Elmer SCIEX Corporation... 

Fig 3. Tryptic Digest of 10 pmol Growth Hormone Releasing Factor... 

Various methods were evaluated for the targeted proteomics of human growth hormone (hGH) in human plasma [111]. hGH was spiked in plasma 10-fold above natural level ( 16 pg/pl). Iiutially, the full plasma proteome was reduced, alkylated, and digested prior to LC-MS via DDA on an ion-trap instrument. hGH could be identified from its T, peptide. Next, the plasma proteome was fractionated by RPLC and GE prior to digestion and LC-MS analysis. hGH could be identified with higher confidence. Finally, an LCxLC-MS approach was apphed, which enabled hGH identification from five tryptic peptides. An important conclusion was that hGH could be detected in a complex sample at the low femtomole level among proteins that were 40,000 x more abundant. The results show that a multidimensional approach may be taken for targeted proteomics and quantitative protein bioanalysis. 

Fig. 1. Elution profile of a tryptic digest of bovine growth hormone on a C,g non-porous silica column, particle diameter 2.1 jam (3.6 cmx8 mm ID). Gradient from 0-50% acetonitrile in 0.1% TFA in 60 min at a flow rate of 1 ml/min.

FIGURE 4,13 Positive ion plasma desorption mass spectrum of the tryptic digest of recombinant human growth hormone after washing with deionized water. (Reprinted with permission from reference 30). 

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