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AF4 Services for AAV Development

Related topics:

Asymmetrical flow field-flow fractionation is an orthogonal method to size exclusion chromatography (SEC) and sedimentation velocity analytical ultracentrifugation (SV-AUC). AF4 is applicable to a wide spectrum of uses, including the detection of soluble aggregates, characterization of species via multiangle laser light scattering (MALLS) and determination of particle size distributions. Follow the link for more information.

Sedimentation velocity analytical ultracentrifugation (SV-AUC) has the potential to become one of the most crucial techniques for evaluation of f adeno-associated virus (AAV) product quality. SV-AUC provides data on the ratio of DNA incorporation into virus capsids. Additionally, this technique gains information about the purity of a product in regard to aggregates and fragments. The importance of these findings makes SV-AUC one of the most powerful stability and quality indicating techniques in the field of gene therapy. Please follow the link for more information.

Dynamic light scattering is a well-established analytical technique in the field of AAV development. Its primary use is to test for aggregate formation. Due to its high sensitivity towards large species, even small impurities caused by aggregation can be detected. We apply our dynamic light scattering instrumentation to determine particle size distributions. Additionally, we use this technique to provide information on thermal stability. Please follow the link for more information.

Size exclusion chromatography (SEC) is the most commonly used technique for aggregate determination in biopharmaceutical development. Obvious advantages are the easy handling and the good reproducibility that this method provides. However, with the recent interest in gene therapy-based APIs the limitations of the technique must be considered. SEC features a limited size range toward larger species, depending on the column, and therefore poses a challenge when a specific method for AAV detection is developed. We are able to tackle that issue, using our experience and state-of-the-art analytical equipment. Please follow the link for more information.

With the increasing interest in gene therapy products also the interested in AF4 is growing. However, AF4 methods are comparably complex to develop. While most people use AF4 only as separation technique. We are experts in AF4 theory and use the theory to get more information out of the technique (e.g. particle size distributions). If you want to get more information about AF4 theory please follow this link.

The detection of soluble aggregates is a crucial step for a wide range of bio-pharmaceutical products. It can be seen as one of the most important quality parameter, independent of the type of material you want us to analyze for you.  if you have a gene therapy, a proteinaceous or an oligonucleotide based product. We can offer a   We are offering  a wide range of analytical techniques for the determination of soluble aggregates. We can support you with finding the best technique to determine the amount of soluble aggregates as well as their characterization. We also have a specialized web page on this very topic. Please follow the link for more information. 

We are offering a wide range of analytical techniques for the determination of particle size distributions.  The detection of nano particle size distributions (1 nm  – 1 μm) is growing in importance. Especially, since the interest in gene therapy increased. Our scientist are working since more than 10 years on the topic of particle size determination. Please follow the link for more information.


General topics:

ZentriForce Pharma was founded as a service provider that specializes in hydrodynamic techniques for biopharmaceutical development. The focus lies on gene therapy products and biosimilarity studies. Follow the link to our company page for further information.

Please follow this link to our contact page

ZentriForce Pharma implemented a strict quality system to assure the highest tractability of results generated at our facility. We have a wide range of quality related standard operation procedures (SOPS) in place to assure the consistency and quality of our laboratory operations. Please follow the link to a description of our quality system. 

ZentriForce Pharma is your expert in analytical services, with focus on hydrodynamic techniques. Please follow the link to find out about our analytical services. 

ZentriForce Pharma offers a wide range of analytical techniques. Please follow the link to our analytical portfolio. 

ZentriForce Pharma is not merely an expert in analytical development. Our scientist are actively involved in further advances in the theory of hydrodynamic techniques. Follow the link to a detailed description of the theory behind our analytical services, including some of our own research

ZentriForce Pharma is always looking for motivated employees. Check out our career page for further information. 

Asymmetrical Flow Field-Flow Fractonation (AF4) Services for Adeno-Associated Virus (AAV) Development

Asymmetrical flow field-flow fractionation (AF4), coupled with a multiangle laser light scattering detector (MALLS), has the potential to develop into a crucial technique for the evaluation of adeno-associated virus (AAV) product purity. It is already one of the main assays within several cases of gene therapy product development.

Our AF4-MALLS instrumentation is optimized for the simultaneous collection of multiple UV signals at highest achievable quality. These measurements can quantify the amount of occurring species and provide information on e.g. molecular weight, UV signal ratio 260 nm/ 280 nm and hydrodynamic radius of each individual species. An additional extraordinary feature of AF4 is its accessible size range. In a single measurement, species ranging from 1 nm up to 1 μm are separated and identified. This remarkable attribute makes AF4 especially valuable for the development of gene therapy products (e.g. Adeno-Associated Virus (AAV) samples)

Please follow the link our AF4 theory page for more details.

AF4 as A Tool in AAV development

Our scientists have worked with a wide range of AAV products. Our experience in AF4 method development keeps your expenses low.

AF4 is a valuable tool for the characterization of AAV samples. One major advantage of the technique is the extensive measurement range (1 nm to 1 μm). Combined with the MALLS signal that gives data on molecular weight and the simultaneous collection of two UV signals (260 nm + 280 nm) extensive characterization of the observed species is possible.  Therefore, AF4 is suitable to analyze both, the contents of a sample and the nature of the   species within (fragments, the AAV and aggregates), in a single run. 

With AF4, we can establish screening methods that cover large measurement ranges and render high resolution data.

ZentriForce Pharma AF4 Instrumentation

Combining the Agilent 1260 Infinity II HPLC, Wyatt DAWN Heleos II MALLS detector and a Wyatt Optilab T-rEX RI detector, we  work with cutting edge instrumentation that offer highest sensitivity,  precision and reproducibility.     

ZentriForce Pharma Exclusive AF4 Services

Our scientists are experts in the application of AF4 to a wide range of bio-pharmaceutical products; further, we continuously and actively work on the advancement of AF4 and the improved utilization of its underlying theory. Starting in 2012, we have developed and improved the AF4Eval program. The software enables us to not only use AF4 underlying theory to determine the particle size distribution of a sample, but also to greatly increase the resolution of the technique by application of a diffusion correction algorithm. We hold the exclusive rights to use the software in a commercial environment but are open to provide our customers with licenses if a method transfer to their facility is requested. The software and its performance is described in detail within the following publication. For more detailed information, follow the link.

Asymmetrical flow field-flow fractonation is an orthogonal method to size exclusion chromatographie (SEC) and sedimentation velocity analytical ultracentrifugation (SV-AUC). AF4 can be used for a wide range of applications ranging from the detection of soluble aggregates, characterization of species via multi angle laser light scattering (MALLS) and determination of particle size distributions. For more information please follow the link.

Sedimentation velocity AUC (SV-AUC) has the potential to become one of the most crucial techniques for the evaluation of the quality of adeno-associated virus (AAV) products. SV-AUC provides data on the ratio of DNA incorporation into virus capsids. Additionally, this technique gains information about the purity of a product, regarding the amount of aggregates and fragments. The importance of these findings makes SV-AUC one of the most powerful stability and quality indicating techniques in the field of gene therapy. Please follow the link for more information.

Dynamic light scattering is a well-established analytical technique in the field of AAV development. It is primarily used to test for aggregate formation. Due to its high sensitivity towards large species, even small impurities of aggregates can be detected within the measurement range of 0.5 nm to 1 µm. Please follow the link for more information.

Size exclusion chromatography (SEC) is the most commonly used technique for aggregate determination in biopharmaceutical development. Obvious advantages are the easy handling and the good reproducibility that this method provides. However, with the recent interest in gene therapy-based APIs the limitations of the technique have to be considered. SEC features a limited size range toward larger species (depending on the column). Therefore, it is challenging to develop a specific method for AAV detection. Nonetheless, our large knowledge base allows us to offer cutting edge analytical method development. Please follow the link for more information.

With the increasing interest in gene therapy products also the interested in AF4 is growing. However, AF4 methods are comparably complex to develop. While most people use AF4 only as separation technique. We are experts in AF4 theory and use the theory to get more information out of the technique (e.g. particle size distributions). We even developed our own AF4 data evaluation software. Please follow the link to more information about our AF4Eval software. If you want to get more information about AF4 theory please follow this link.

Our scientists are not only applying AF4 we are also working on the advancement of AF4 and its theory. Since 2012 we are working on the development of the AF4Eval program. Since then it was continuously improved. The software enables us to not only to use AF4 theory to determine the particle size distribution but also to tremendously increase the resolution of the technique by applying a diffusion correction algorithm. Please follow the link for more information. 

The detection of soluble aggregates is a crucial step for a wide range of bio-pharmaceutical products. It can be seen as one of the most important quality parameter, independent of the type of material you want us to analyze for you.  if you have a gene therapy, a proteinaceous or an oligonucleotide based product. We can offer a   We are offering  a wide range of analytical techniques for the determination of soluble aggregates. We can support you with finding the best technique to determine the amount of soluble aggregates as well as their characterization. We also have a specialized web page on this very topic. Please follow the link for more information. 

We are offering a wide range of analytical techniques for the determination of particle size distributions.  The detection of nano particle size distributions (1 nm  – 1 μm) is growing in importance. Especially, since the interest in gene therapy increased. Our scientist are working since more than 10 years on the topic of particle size determination. Please follow the link for more information.

Interested in AF4 Services ?

We are more than happy to assist you with our AF4 services.

Asymmetrical Flow Field-Flow Fractionation Specificiation

Main OutputSize distribution (rH or dH); relative concentrations; molecular weight (Mw)
Size range1 – 1000 nm
Additional Outputsradius of gyration (rg) 
Sample Volume4 µL to 100 µL
Measurement typeHPLC vial based
Temperature range 18°C to 40°C
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Asymmetrical flow field-flow fractonation is an orthogonal method to size exclusion chromatographie (SEC) and sedimentation velocity analytical ultracentrifugation (SV-AUC). AF4 can be used for a wide range of applications ranging from the detection of soluble aggregates, characterization of species via multi angle laser light scattering (MALLS) and determination of particle size distributions. For more information please follow the link.

Sedimentation velocity AUC (SV-AUC) has the potential to become one of the most crucial techniques for the evaluation of the quality of adeno-associated virus (AAV) products. SV-AUC provides data on the ratio of DNA incorporation into virus capsids. Additionally, this technique gains information about the purity of a product, regarding the amount of aggregates and fragments. The importance of these findings makes SV-AUC one of the most powerful stability and quality indicating techniques in the field of gene therapy. Please follow the link for more information.

Dynamic light scattering is a well-established analytical technique in the field of AAV development. It is primarily used to test for aggregate formation. Due to its high sensitivity towards large species, even small impurities of aggregates can be detected within the measurement range of 0.5 nm to 1 µm. Please follow the link for more information.

Size exclusion chromatography (SEC) is the most commonly used technique for aggregate determination in biopharmaceutical development. Obvious advantages are the easy handling and the good reproducibility that this method provides. However, with the recent interest in gene therapy-based APIs the limitations of the technique have to be considered. SEC features a limited size range toward larger species (depending on the column). Therefore, it is challenging to develop a specific method for AAV detection. Nonetheless, our large knowledge base allows us to offer cutting edge analytical method development. Please follow the link for more information.

With the increasing interest in gene therapy products also the interested in AF4 is growing. However, AF4 methods are comparably complex to develop. While most people use AF4 only as separation technique. We are experts in AF4 theory and use the theory to get more information out of the technique (e.g. particle size distributions). We even developed our own AF4 data evaluation software. Please follow the link to more information about our AF4Eval software. If you want to get more information about AF4 theory please follow this link.

Our scientists are not only applying AF4 we are also working on the advancement of AF4 and its theory. Since 2012 we are working on the development of the AF4Eval program. Since then it was continuously improved. The software enables us to not only to use AF4 theory to determine the particle size distribution but also to tremendously increase the resolution of the technique by applying a diffusion correction algorithm. Please follow the link for more information. 

The detection of soluble aggregates is a crucial step for a wide range of bio-pharmaceutical products. It can be seen as one of the most important quality parameter, independent of the type of material you want us to analyze for you.  if you have a gene therapy, a proteinaceous or an oligonucleotide based product. We can offer a   We are offering  a wide range of analytical techniques for the determination of soluble aggregates. We can support you with finding the best technique to determine the amount of soluble aggregates as well as their characterization. We also have a specialized web page on this very topic. Please follow the link for more information. 

We are offering a wide range of analytical techniques for the determination of particle size distributions.  The detection of nano particle size distributions (1 nm  – 1 μm) is growing in importance. Especially, since the interest in gene therapy increased. Our scientist are working since more than 10 years on the topic of particle size determination. Please follow the link for more information.