AAV Affinity Chromatography Resins (Type II)

AAV Affinity Chromatography Resins (Type II) are used for Rapid and Efficient Purification of Adeno-Associated Virus (AAVII). The chromatographic operation typically includes equilibrium, sample loading, washing, elution, and cleaning steps. The specific operating methods are as follows:

Equilibrium: Use 5-10 column volumes (CV) of equilibrium buffer (AAV binds to AAV affinity chromatography medium near neutral pH, so conventional buffer systems such as PBS, Tris, and citrate can be used for AAV purification. For example, 20 mM PB + 0.5M NaCl, pH 7.4) to equilibrate the ion exchange column at a flow rate not exceeding the maximum pressure limit, until the conductivity and pH of the effluent remain constant (consistent with the equilibrium buffer).

Sample Loading: The sample buffer should be as consistent as possible with the equilibrium buffer. To avoid clogging the chromatography column, the sample solution should be centrifuged or microfiltered (0.45μm). The sample loading volume should be calculated based on the resins' binding capacity and the AAV content in the sample.

Washing: After loading, continue washing with the equilibrium buffer until the UV absorbance drops to baseline.

Elution: Use elution buffer (usually by reducing pH to elute, e.g., within the range of pH 2 to 5, such as 0.1 M glycine + 0.5M NaCl, pH 2.5) to collect the effluent. Since AAV is sensitive to highly acidic conditions, the collected eluate should be neutralized immediately.

Cleaning: After use, the AAV affinity chromatography resins need to be cleaned (nanobodies have low alkaline stability, so NaOH solution is not recommended for AAV affinity chromatography resins cleaning). The recommended cleaning steps are as follows:(1) Wash the column with 0.1M citric acid pH 2.0 (or 0.1M glycine pH 2.0);(2) Wash the column with 2M guanidine hydrochloride or 2M urea;(3) Re-equilibrate with neutral pH buffer (such as the equilibrium buffer 20 mM PB + 0.5M NaCl, pH 7.4) for the next purification cycle, or store in 20% ethanol.

Storage: Store at 2-10°C in 20% ethanol; the unpacked resins can be flushed with 20% ethanol and stored at 2-10°C.

Other considerations: During use and storage of the column, avoid running dry or improper sealing to prevent air bubbles from entering.

3.Specification