LOW SURFACE TENSION
BioPharm: Low surface tension of some adjuvant solutions can reduce the efficiency of filters' bacterial retention. How can this problem
Protein Purification Using Single-Use Technology
Bromm (Sartorius Stedim): It is advisable and required by regulators to carry out a comprehensive filter validation study, including bacteria-retention
testing, simulating worst-case process parameters with actual product formulation using process related (i.e., pleated) scale-down
filter devices. The design of the filtration system should consider reducing filtration time and differential pressure because
these two parameters, among others, may increase the risk for bacterial breakthrough. During a filter evaluation study, the
impact of different inlet pressure filtration conditions should be assessed, including constant flow or constant pressure
conditions. Constant flow conditions may increase the risk of bacterial breakthrough, because of the increased differential
pressure required to keep the flow constant during the filtration process and increased filter blocking.
The use of filters specifically designed for adjuvant filtration as explained above is highly recommended because those filters
will keep the process parameters at a moderate level. It is recommended to carry out a bacterial-retention study early in
the filter-selection process to find the optimal solution based on retention efficiency and highest filtration capacity.
Martin (Pall): Statistical and empirical studies at Pall Corporation have identified low surface tension of some adjuvant solutions as a
risk factor for reduced bacterial retention efficiency of most sterilizing grade 0.2 μm rated membrane filters. The mechanism
by which bacterial retention is reduced under lower surface tension in these fluids is not yet fully elucidated. Some mitigating
factors appear to be membrane structure and layering of multilayer media, operating conditions, as well as reduction of bacterial
bioburden or challenge levels and reducing challenge duration. Fluid surface tension affects the interactions between the
bacteria and the membrane flow-path surfaces, but detailed mechanisms are not well known and specific surface tension thresholds
cannot be determined.
Membrane surface chemistry is also an element that may mitigate the negative impact of fluid surface tension. Determining
how and to what extent membrane-surface chemistry can enhance retention requires extensive studies. Filters with positive
zeta potential, which provide enhanced adsorptive removal properties for bacteria in aqueous ionic solutions, have been used
in the past for such purposes. This was also one of the capability advantages of asbestos-containing filters, although these
are no longer used because of asbestos safety concerns.
Koklitis (3M): Such reduced filter efficiency can be related to the mechanisms involved in bacterial retention, which can be based not only
on sieving but also on entrapment and electrostatic attraction. The adsorption of bacteria to the membrane polymer surface
can be caused by any combination of forces, including hydrogen bonding, charge-induced, and Van der Waals interactions. The
presence of liposomes, oils, or surfactants in a process stream can disrupt these adsorptive interactions and consequently
reduce retention of bacteria within the membrane structure.
When there may be a high risk of bacterial penetration, it should be identified and considered in the planning of a filter
validation study. The required minimum bacteria challenge (1 × 107 colony forming units of Brevundimonas diminuta per cm2 effective filter area) must apply, although an upper challenge level can be considered and restricted to one log higher.
In a full-scale production process, the bacterial challenge to the final filter membrane may be controlled by introducing
a prefiltration stage that has been demonstrated to be effective for bioburden reduction. The careful management and control
of the operating conditions during process filtration will also help mitigate the risk of bacterial penetration, with attention
to flow rate and filter area sizing to avoid high pressure drop.
Powell (Asahi): This issue is typically not applicable to Asahi products, but with some filters, the lower surface tension can change the
effective porosity rating of the membrane, allowing larger particles to slip through the membrane's holes. These low viscosity
adjuvants effect the thickness of the boundary layer (where flow velocities at the membrane surface are at or close to zero)
which, in turn, alters the effective pore size under those conditions. It can also affect how the API and contaminants build
up around the membrane's pores hence altering the effective pore size. One can screen different membrane types, porosities,
and brands of filters, and work closely with the membrane filter supplier to choose the best filter for the application.
BioPharm: Can certain types of adjuvants cause fewer problems with regard to filters' bacterial retention?
Bromm (Sartorius Stedim): A review of validation studies and field tests for a broader variety of fluid formulations indicates that low surface tension
formulations, such as many adjuvants or adjuvanted vaccines, present a higher risk for bacterial penetration of sterilizing-grade
membrane filters. Among such formulations, according to the data analyzed, liposome formulations present a higher risk than
surfactant containing solutions. Therefore, the use of such formulations may be a suitable alternative to replace more critical
formulations where applicable.
Martin (Pall): It is possible that certain adjuvants and related low surface tension fluids may be intrinsically less likely than others
to cause reduced retention efficiency by membrane filters. However, there is insufficient data at this time to draw firm conclusions
and make recommendations. In addition, awareness among vaccine producers that selection of surfactant-containing adjuvants
and processing conditions can influence bacterial retention efficiency of sterilizing filters is not yet widespread. Until
then, filter manufacturers must continue to work with vaccine developers to define appropriate membranes and optimize reasonable
processing conditions to sterilize any vaccine formulation. Certainly, elaboration of an optimum adjuvant with such a goal
would require an extensive amount of work and a very close partnership between filter manufacturers and vaccine producers.
Koklitis (3M): The choice of adjuvant is dependent on meeting the requirements of the process under consideration. The pros and cons of using
a particular type of adjuvant must be considered and compared. When liposomes are selected as adjuvants their role as antigen
carriers is utilized along with their immunological enhancement effect.
Powell (Asahi): These issues should be discussed with the membrane supplier's technical support teams and if they can't help, the filters
must be screened to choose the best solution for the filter application. The answer depends on the membrane chemistry, but
for large porosity filters, surfactant-containing solutions are typically not that large of a problem. Smaller porosity filters
can be dramatically impacted in a negative way.