There is increasing pressure, even by regulatory agencies, like the European Commission’s Joint Research Centre, to avoid and even ban generation and use of animal-derived antibodies, according to the 3R's principle on animal research and the existence of alternative methods such as phage display technology. However, several key question seem unclear:
- Does non-animal-derived antibodies have similar affinity and specificity than animal-derived ones? Does they work fine in all applications?
- Is there enough scientific evidence supporting a quick shift toward non-animal-derived antibodies, including banning of those generated in animals?
- Currently, can phage display technology successfully replace hybridoma technology?
Comments on this issue from researchers with experience on use of non-animal-derived antibodies are welcome, especially those from immunologists.
In fact large quantities of monoclonal antibodies for therapeutic use are currently produced at industrial scale by genetically modified cells (not in animals). So the antibody production field is mostly animal-free already.
But the original source of variable regions for such recombinant antibodies can be either hybridomas from immunized animals or libraries displayed on phage, yeast...and even a combination of both- libraries made using lymphocytes of immunized animals. Antibodies for research or analytical purposes are also frequently produced using hybridomas.
You can find published examples showing superior quality of the antibodies coming from in vivo immunization, as well as other papers showing extremely high affinity and specificity of library-derived antibodies without any immunization step.
As a scientist, I think the ideal scenario would be to have both alternatives to choose the best in every case according to the particular application and the expertise of the lab and people involved. I have experience working with both hybridomas and phage-displayed libraries.
But in case of an eventual replacement of animals since the initial steps of antibody development by in vitro sources, libraries have been shown to produce antibody fragments that can be converted to whole antibodies having high affinities and the desired specificity properties, even when working with challenging targets. So I think fully in vitro antibodies could do the job very well if antibody-producing companies use libraries of enough size and diversity, high throughput selection methods to pick the ideal binders, and careful engineering strategies to further improve them if necessary. In fact many companies are doing this already.
In the case of smaller labs that make excellent antibodies for particular applications, the situation is different, because some of them do not have expertise in display techniques or their libraries and selection platforms are not so powerful to produce any desired binding site. For them an eventual transit to animal-free antibody generation could be slow and difficult, and the scientific community could loose many valuable antibodies in the meantime. I think that a growing bias towards the use of antibodies from non-animal sources (starting with big companies) would be much better than a ban.
I'm afraid this is in fact unlcear. You can find papers where natural antibodies have superior KD. You can also find papers where phage-displayed ones do. But what you want, to base a policy decision on, is a wide-ranging comparison across many target antigens and different applications, done by a group that's credibly disinterested in the outcome. I don't believe this exists.
In fact large quantities of monoclonal antibodies for therapeutic use are currently produced at industrial scale by genetically modified cells (not in animals). So the antibody production field is mostly animal-free already.
But the original source of variable regions for such recombinant antibodies can be either hybridomas from immunized animals or libraries displayed on phage, yeast...and even a combination of both- libraries made using lymphocytes of immunized animals. Antibodies for research or analytical purposes are also frequently produced using hybridomas.
You can find published examples showing superior quality of the antibodies coming from in vivo immunization, as well as other papers showing extremely high affinity and specificity of library-derived antibodies without any immunization step.
As a scientist, I think the ideal scenario would be to have both alternatives to choose the best in every case according to the particular application and the expertise of the lab and people involved. I have experience working with both hybridomas and phage-displayed libraries.
But in case of an eventual replacement of animals since the initial steps of antibody development by in vitro sources, libraries have been shown to produce antibody fragments that can be converted to whole antibodies having high affinities and the desired specificity properties, even when working with challenging targets. So I think fully in vitro antibodies could do the job very well if antibody-producing companies use libraries of enough size and diversity, high throughput selection methods to pick the ideal binders, and careful engineering strategies to further improve them if necessary. In fact many companies are doing this already.
In the case of smaller labs that make excellent antibodies for particular applications, the situation is different, because some of them do not have expertise in display techniques or their libraries and selection platforms are not so powerful to produce any desired binding site. For them an eventual transit to animal-free antibody generation could be slow and difficult, and the scientific community could loose many valuable antibodies in the meantime. I think that a growing bias towards the use of antibodies from non-animal sources (starting with big companies) would be much better than a ban.
Absolutely agree with Gertrudis Rojas , the affinity of antibodies generated in vivo versus the ones generated through phage display widely depends on the way they were generated.
I would just like to share a recent interview with a researcher with expertise in using this technology for antibody discovery:
https://www.proteogenix.science/scientific-corner/antibody-production/cell-based-phage-display-biologically-relevant-antibodies/
In this interview, he explains the advantages and limitations of the technology and discusses how the field is likely to evolve in the foreseeable future.
It is true that phage display-derived antibodies can sometimes exhibit lower affinities and higher aggregation profiles, however, there are already a lot of techniques in place that allow us to bypass these constraints, namely:
- In vitro affinity maturation using 3D modeling to construct high-quality mutant libraries that can subsequently be screened with phage display
- The use of bioinformatics tools to determine potential developability issues (low stability, high aggregation, etc.) as early as possible
- Subjecting in vitro generated antibodies to high temperatures to ensure their long-term stability
- Using the antigens in their native or near-native conformations to obtain as many biologically relevant antibodies as possible (i.e. using cell-based phage display for membrane-bound antigens)
Moreover, phage display is particularly useful for generating antibodies against toxic and non-immunogenic antigens, which currently cannot be obtained through hybridoma technologies.
And due to the simplicity of this approach, it is possible to tailor the cross-reactivity of an antibody very easily which is useful in two situations:
https://www.proteogenix.science/scientific-corner/antibody-production/how-advanced-monoclonal-antibody-generation-methods-are-improving-conventional-envenoming-treatments/
Hope this information was helpful!
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