Olignucleotide Molecular Weight Analysis
Oligonucleotide molecular weight determination among other biopolymers can be determined at the mass spectrometry facility in IMSERC. An Agilent 6220A Time-of-Flight LCMS system (resolving power of ~10,000) is used to perform an online sample clean up by liquid chromatography followed by electrospray ionization, high resolution mass spectrometric analysis. The complexity of the raw data (due to significant contribution from adjacent molecular isotopes under each one of the charge state ion envelope) makes it nearly impossible to manually interpret the data. Maximum Entropy algorithm is utilized to deconvolute the data into a simple neutral spectrum where each unique molecule is assigned a single Mr. Relatively small biopolymers of up to 150 kDa has been successfully analyzed.
Below are spectra of raw (left), deconvoluted data (middle), and a zoomed inm spectrum showing potential modifications to the specific biopolymer.
- Determine the molecular weight of an oligonucleotide of interest.
- Detect impurities in olignucleotide synthesis. These could be omitted bases, truncations, or other reaction biproducts.
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