Pronalyse is a protein characterization division of Creative Proteomics, an integrated CRO with rich experience in providing drug development services. Nucleotide-based drugs are well established and represent an important class of clinical agents. Although these drugs are highly effective, the synthesis and development of these molecules are susceptible to process-related impurities and modifications, and complete characterization is important throughout the development and production process. To this end, Creative Proteomics Pronalyse has introduced highly sophisticated oligonucleotide characterization services.
The potential of therapeutic oligonucleotides as an emerging class of therapeutic modalities continues to be explored and is leading a global wave of research and development. However, oligonucleotides are much larger than traditional small molecules, and their preparation involves a large number of synthetic steps, with typical multi-step cycles of solid-phase synthesis likely to introduce impurities/invalid sequences, making analysis and characterization more difficult. In addition, oligonucleotides have certain specific physicochemical properties, such as: usually large negative charges; PO impurities from phosphorylation; hydrolysis of purine bases to form depurine fragments; and hydroxyl groups on ribose, which are difficult to separate from the main and full-length products, so the development of oligonucleotides usually relies heavily on mass spectrometry analysis and quantification.
Oligonucleotides are usually composed of 20 or fewer short-stranded nucleotides (deoxyribonucleotides or ribonucleotides) that can pair with DNA, mRNA or pre-mRNA to achieve a very high degree of selectivity through the Watson-Crick principle of base complementary pairing, precisely inhibiting certain genes and silencing genes that encode abnormalities, thus preventing many mis-expressed proteins. Many types of oligonucleotides are currently under development, including antisense oligonucleotide (ASO), small interfering RNA (siRNA), microRNA (miRNA), and nucleic acid adaptor (aptamer).
“Oligonucleotides (ONs) are becoming increasingly important as promising new biologic drugs, and Creative Proteomics’ advanced mass spectrometry platforms and analytical systems offer an efficient and convenient option for impurity analysis. Inspired by protein biomolecule mass spectrometry, we are able to use tandem mass spectrometry to identify long oligonucleotide fragment sequences,” commented Pronalyse’s project manager.
Creative Proteomics Pronalyse uses MALDI-TOF or ESI-MS mass spectrometry to characterize and identify oligonucleotide impurities, and each technique reserved its advantages and disadvantages. Pronalyse applies one or both instruments for your analysis, depending on the synthesized sequence’s nature.
1. MALDI-TOF MS
MALDI-TOF MS is well suited for high throughput requirements and characterization of nucleotides up to 50 bases in length.
2. ESI-MS
ESI-MS is most suitable for oligonucleotide applications larger than 50 bases (>13 kDa), such as gene synthesis, and has upregulated instrumental accuracy for molecule descriptions.
About Creative Proteomics Pronalyse
Pronalyse is supported by specialists who are professional and skilled in protein chemistry, NMR, HPLC, mass spectrometry and bioinformatics. Currently, Pronalyse is dedicated to providing protein-oriented analysis services to support manufacturers and researchers in biopharmaceutical development, mainly including antibodies, PEGylated proteins, peptide and protein drugs, vaccines, amino acids and their derivatives. The services provided include in vitro characterization, purity analysis, protein structure modification, in vivo safety checks, and pharmacokinetics analysis, etc.
