Reference Library-
Case Studies

Situation

The customer has developed a new vaccine or other molecule in the research laboratory. In the course of research and development of the product, the ‘gold standard’ used for particle separation was ultracentrifugation. Now the plan is to start pilot plant studies with the final target of full scale manufacturing.

Objectives

• Reproduce the results obtained in the laboratory

• Scale up the volume without modifying the results

• Obtain the highest recovery levels

• Minimize the quantity of consumables handled and disposed of

• Retain the infectivity of the original molecule

• Operate in an aseptic environment

AWST Response

The Continuous Flow Ultracentrifuge mimics the activity of the vertical rotor in the lab.

AWST’s applications specialists will assist in transferring the separation protocol developed in a vertical rotor in a lab ultracentrifuge to the use in our continuous flow ultracentrifuge. The AW Promatix 1000™ BioPurification System enables the initial methodologies to be established that can then be transferred directly to pilot scale and production scale ultracentrifuges. This avoids the complex work needed to transpose from results obtained in an ordinary laboratory ultracentrifuge.

Scale-up is achieved with no modification whatsoever to separation results.

The use of the PKII ultracentrifuge plus PK3 rotor with 200mL core enables starting volumes of about a litre to be used for replicating the results and proving the efficiency of transfer from the lab ultracentrifuge.

As pilot scale production evolves, the rotor core alone has to be replaced. The possibilities are 400mL, 800mL and 1.6 Litre rotor volume with working volumes attaining over 100 Litres. Exactly the same path length exists in every configuration so exactly the same protocol parameters are applied in every case. If desired, a KII centrifuge can be introduced with operational processing of over 200 Litres.

Very high recovery levels are maintained by the single step procedure.

If necessary, a PK6 rotor can be used. This includes a pre-clarifier for larger volumes of fluids with a high load of unwanted particles. Thus Clarification, Concentration and Purification are carried out in one step. This avoids loss of product through multiple steps.

The number of consumable items is low and the items are small and easy to handle.

No awkward filters, columns or buffers to dispose of; just small seals and maybe some tubing.

Infectivity is retained at the original level.

Low shear forces are applied throughout the process with the particles always suspended in fluid as protection against disruption. Negligible damage to the particle means that activity is maintained.

Aseptic ultracentrifuge available.

The new aseptic ultracentrifuge enables operators to ensure total protection for the product throughout the production procedure. The use of the AlfaSIP™ sanitization kit introduces a major advantage in the aseptic operation of the ultracentrifuge.

Reference Library- Case Studies