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
 
 

HOME
 
 

Date Last Modified:  17 APRIL 2012