The capsid of adeno-associated viruses (AAVs) consists of three viral proteins (VPs, 62-90 kDa). Alterations in the dedicated protein composition can influence the vector's potency and increase the risk of immunogenicity.

To reliably determine the ratio of the three proteins VP1, VP2 and VP3, reversed phase liquid chromatography (RP-LC) coupled to mass spectrometry (MS) is a viable option. Only in combination with bioinert column hardware can a good separation of all three proteins be achieved. When using stainless steel column hardware, the negatively charged capsid surface interacts with the column body, which results in a decrease in resolution.

The advantage of using bioinert YMC-Accura Triart Bio C4 columns when analysing AAV capsid proteins: sharp peaks and excellent resolution.

This Technical Note demonstrates the advantage of using a bioinert YMC-Accura Triart Bio C4 column for the analysis of AAV capsid proteins.

For more information on YMC-Accura Triart columns please look here or check out the YMC-Accura Triart flyer.

IP-RP is commonly used for the analysis of oligonucleotides. Due to their highly polar nature HILIC can be an alternative approach. This Expert Tip compares the analysis of oligonucleotides using both modes and demonstrates the preference for each mode.

In both modes the evaluated poly(dT) DNAs were separated by length starting with the shortest. Furthermore, DNA and RNA with the same sequence show similar retention in HILIC mode, only the DNA is more strongly retained in IP-RP mode. Different chemical RNA modifications, fully phosphorothioated and fully methoxylated showed different separation behaviour in HILIC and IP-RP.

For full details read the Expert Tip.

Please visit our homepage or download the YMC Biochromatography columns catalogue to get further information about YMC’s BioLC solutions.