YMC i-Mail August 2023

YMC-Accura Triart Bio C4

The best choice for AAV capsid protein analysis

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.

Figure: Comparison of AAV capsid VPs separation RP-LC using the bioinert YMC Accura Triart Bio C4 and another C4 column with stainless steel hardware.

New brochure

New YMC brochure on oligonucleotide columns with lots of helpful information

  • Applications for a variety of different oligonucleotides
  • Column selection for IP-RP, AEX, SEC or HILIC mode
  • Detailed information about each separation mode and benefits
  • Several expert tips for affecting the separations for the different separation modes



Expert tip

When HILIC is a suitable alternative to IP-RP for the analysis of oligonucleotides

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.

Figure: Analysis of RNA (top), phosphorothioated RNA (middle) and methoxylated RNA (bottom) with the same sequence using HILIC (left) and IP-RP mode (right).