PROTEINS,
PEPTIDES
&
OLIGONUCLEOTIDES
ROCKY MOUNTAIN INSTRUMENTAL LABORATORIES
RMIL provides cGLP (FDA quality) analysis of proteins, peptides, oligonucleotides, and traditional pharmaceutical molecules in raw materials, finished products and clinical biosamples.
BIOPOLYMERS,
such as proteins, peptides, polysaccharides, and oligonucleotides, can
be analyzed in raw materials, formulated products, and in clinical biosamples,
such as serum, plasma, and solid tissues. As with any analysis, the
emphasis should be on the data needed, rather than on the technology used
to obtain that data.
For
example, formulated protein pharmaceuticals may have significantly altered
secondary and tertiary structure, which may be detected by the use of circular
dichroism (CD) or FT/IR analysis of liquids or solids (1). If deamidation,
iso-aspartate formation, or other post-translational modification
may be a problem, this is probably most efficiently investigated by the
use of HPLC/MS/MS (2, 3), although other
MS techniques are often useful. Similarly, phosphorylation
and glucuronide formation can be detected using HPLC/MS/MS. In particular,
it is possible to use a tandem MS/MS to detect those molecules which contain,
specifically, phosphorylation, glucuronidation, or other post-translational
modification with excellent overall sensitivity because only those molecules
will be detected by the MS detector.
Oligonucleotides
can be analyzed in raw material or formulated product by the use of MALDI/TOF,
ESI/Quadrupole-TOF, ICRMS, HPLC/MS, or HPLC/MS/MS. Structures, including
sequences, are confirmable based upon the mass spectra of the products.
In particular, PS failures and incorrect nucleotide sequences are easily
detectable. Similarly, modified bases can be detected and identified. HPLC/MS/MS
also is quite useful for the analysis of synthetic oligonucleotides and
metabolites in biological samples (4).
If you have a need for structure determination using either FT/IR or LC/MS/MS, please contact us. We have significant experience in the analysis of pharmaceuticals as well as synthetic and natural polymers. Also see the links on our home page.
Of particular interest may be the EMBL, Protana, NewObjective, and ASMS sites and links therein.
REFERENCES:
1. Ahern, T. and M. Manning "Stability of Protein Pharmaceuticals" Part A, Plenum Press, 1992.
2. Chapman, J. "Protein and Peptide Analysis by Mass Spectrometry" Humana Press 1996. Excellent information on exactly how to do LC/MS of biopolymers.
3. Marshak, D. (ed.) "Techniques in Protein Chemistry" Academic Press. This is a series, based upon the annual meeting of the Protein Society. Excellent detail.
4. Nordhoff, E. et al "Mass Spectrometry of Nucleic Acids" Mass Spec. Rev., 15, pp 67-138 (1996).
5. Lantz, R. K. and Sulik, P.L. "HPLC/MS: A User's Perspective" European Pharmaceutical Contractor, October, (1998).
6. Walker, John (ed.) "The Protein Protocols Handbook" Humana Press (1966) Very practical compilation of 144 methods for all sorts for general protein and peptide work. Specifically avoids instrument intensive methods, such as HPLC/MS or NMR. Spectacularly useful.
7. Vervoort, R.J. M. et al "Preliminary Study on the effect of miniaturization and use of volatile mobile phases in LC for the on-line LC-MS analysis of basic pharmaceuticals." J. Pharm. Biomedical Analysis 21 273-289 (1999).
8. Cole, R. B., ed. "Electrospray Ionization Mass Spectrometry" J. Wiley (1997). Excellent text on ESI MS, with applications and useful reviews.
All of the above books are available from B&N or Amazon, for which there are links on our home page.
There are many
excellent web sites for proteomics work. One which has many useful links
is Biobase (Denmark) and
the National Center for Biotechnology Information (NCBI).
Also
see the many links on our
home page.
Robert K. Lantz, Ph.D., Director.
970-266-8108 303-530-1169
Rocky Mountain Instrumental Laboratories, Inc.
108 Coronado CT.
Ft. Collins,
CO 80525
Date Last Modified:
17 APRIL 2012