Focus Areas / Animal-Free Products

Genetic Manipulation and drug testing have extensively involved the use of animals in one way or the other. In the UK, more than three million animals are used for research purposes despite the availability of alternative methods of experiementation such as in-vitro approaches using DNA library phage technology.

The concerns around quality, inconsistency of the formulation, lack of validation of reagents used in research as well as poor reproducibility of results using animal derived approaches have been major concerns in the bioscience community. More importantly, researchers must bear costs of animal usage as it requires breeding, housing, feeding, transporting, and disposing of animals. By using animal-free products, researchers or manufacturers can can avoid these ethical and practical issues, thereby focus on more humane and efficient reserach approaches.

By using animal-free products, researchers can reduce their reliance on animals, improve their research quality and relevance, and accelerate the translation of their findings to human health.

Another advantage of animal-free products is that they improve the quality and relevance as well as efficiency of research results. Animal models are often poor predictors of human responses, as they have different physiology, genetics, metabolism, and immune systems. This leads to inaccurate and unreliable data that may not be applicable to humans due to immunogenicity differences and genetic drifts. For example, a study by Harvard University in 2020, found that mouse models failed to mimic human inflammatory diseases (5).

With these numerous concerns, there has been a collective effort in the research community to adopt human-based / animal free in-vitro assays to replace animal models where possible and increase the predictivity of preclinical research (2).

One of the major shifts has been the synthesis of recombinant antibodies by manufacturers, where usage of different expression vectors from prokaryotic to eukaryotic offering diverse choices for the researchers. The expression systems include E. coli, Yeast, Baculovirus infected insect cells, Mammalian Cells, and in-vitro. E. coli that can encode the DNA of interest for a selected antibody chain. The expression vector can be transfected into a mammalian cell lines such as HEK or CHO which allow production of engineered antibodies (see Figure 1) (1).

Figure 1 - Methodology of Manufacturing Recombinant Antibodies

Figure 2 - Methodology of obtaining DNA for production of recombinant antibodies

Several organisations and initiatives like the National Centre for Replacement Refinement and Reduction of Animal in Research (NC3Rs), the European Union Reference Laboratory for Alternatives to Animal Testing (ECVAM), are trying to initiate the relevance of the 3Rs concept established by Charles Hume and William Russel in 1959 that states (4):

Reduction as the decrease in the number of animals in experimentation by using only the required number of animals for procedures.
Refinement for the improvement of procedures to reduce invasiveness and animal stress.
Replacement for the search of alternatives to animal experimentation and animal-derived products use in research.

Recently in 2020, the EURL ECVAM Scientific Advisory Committee advised against animals being used for the development and production of antibodies and made several recommendations of promoting use of non-animal derived antibodies going further. The report from the EU derivatives also proposes that, EU should refuse grant funds to projects using only animal derived antibodies (1). Also, in Europe the testing of cosmetics products on animals was banned in 2004, and later, in 2009, the regulatory measures were updated to demand that cosmetic ingrediants' safety must be assessed by non-animal methods.

These initiatives are slowly making their was within the research environment with manufacturers looking for new approaches to enhance their reagents capability in recent years.

A small study by Pivotal Scientific Limited, have offered great insights about the approach manufacturers and distributors are taking in relation to the European Union Directives. As per the survey response it has been seen that:

60% of suppliers and 65% of distributors mentioned they are already selling non-animal derived antibodies (1).
10% of the distributors noticed a rise in demand for these products and 22.5% saw an increase in supply over the past years (1)

According to Marlies Halder, co-author of the EURL ECVAM Recommendation "Animal - derived antibodies typically suffer from batch-to-batch variability and show low specificity to targeted molecules. These problems can be easily solved using non-amimal derived phage display technology. Their use will improve reproducibility, and relevance of scientific procedures and lead to more efficient and effective use of research funds" (3).

Use of animal free products can improve research capability. Some of the other benefits of non-animal derived reagents are:

	Benefits of Non-Animal derived Reagents	Features of Non-Animal derived Reagents
Specificity	Customisation of reagents can provide efficient results, reproducibility, reduce costs and allow more time to perform multiple research approaches.	Developed in a phage library so design and conformation are already known.
Faster Production	More time for research management and planning, including less waiting time.	Process can occur in a week rather than 3 to 4 months.
Supply	Instead of a one-off experiment, multiple research approaches are possible because of the high production capability.	Larger production and availability
Contamination	Genetic drift and batch variability are controlled; therefore, results are reproducible and validated with duplicated experiments.	No contamination as non-animal technologies are of human origin and no batch-to-batch variability since protocols are optimised and controlled
Costs	Helps to cut costs and allows more funding opportunities	With high level of specificity through customisation and phage technology, including large production capability, the costs can offset those of polyclonal antibodies
Applicability	Customised and lab-based approach helps to manage the research timelines	Broader application as it is a bench-based approach and easily reproducible

2BScientific is a leading distributor of life science products in the UK and Europe. Our mission is to provide high-quality and innovative products that meet the needs of researchers in various fields of science. We believe that animal-free products are not only ethical, but also reliable in terms of quality and relevance. That is why we work towards supporting our suppliers on their sustainability initiatives that involve the production of animal-free products. Therefore, 2BScientific offers a broad range of animal free products that include:

Recombinant Antibodies and Proteins
Animal free Neural cell culture mediums and serum
Animal free buffers
Animal free Cytokines
Recombinant Lectins for Bio-affinity proteins
Lectin Binding Assays

Recombinant Antibody and Proteins

Absolute Biotech Company that includes Absolute Antibody and LSBio, have both been extensively involved in being part of the Animal Free antibody movement, and have developed alternatives to offering Recombinant Antibodies and Proteins that require no immunisation of animals. The platform is based on transient, chemical transfection of HEK293 cell lines cultured in a serum free medium. No viral transfection approaches are used.

The platform can also be used for Custom Antibody Expression Services and is used to produce the rapidly growing Recombinant Antibody and Protein Catalog.

Key Benefits

Through their recombinant technology they ensure reproducibility, defined conformation knowing what is in the vial, high purity as they use Serum-free mammalian expression system, and a large variety of antibody formats can be produced.

Absolute Antibody Reagents LSBio Animal-Free Proteins

Cape Biologix Technologies uses plant-based expression platforms to develop recombinant proteins. A unique alternative where they sequence the DNA digitally and then synthesise it into a physical DNA by using a high expressing PCBP plant expression vectors. The expression vectors are then inserted into Agrobacterium Bacterium. Plants are infiltrated with a solution of transformed bacteria, allowing the delivery of vectors into the cells of plants. A week after infiltration, the leaves of the infected plants are harvest for protein extraction and purification, further they are subjected to stringent quality testing to ensure purity, functionality, and performance.

Key Benefits
Using these expression systems for recombinant proteins, they provide wider application capability such as Cell assay, Binding assay/Protein-protein interaction, Cell culture-serum-free medium, Drug related studies, Enzymatic activity in vitro, ELISA standard and its raw materials, In vivo study, Mass spectrometry standard, Protein chip, SDS-PAGE control, Protein structure analysis (Crystal/ electron microscope). Also, to enhance results analysis and research efficiency, they provide multiple tags that can be customised as per request.

Animal Free Diluent and Blocker Buffers

Kementec and Vector Laboratories offers animal free buffers to improve assay sensitivity, reduce non-specific binding and ensure no cross reactivity with animal sourced antigens and antibodies. These buffers are BSA free thereby reduces regulatory concerns and offers faster cross border availability.

Animal Free Neural Cell Cultures Mediums

Creative Biolabs provides Animal free culture medium for ~~the~~ cell culture, maintenance, and differentiation of pluripotent and Embryonic stem cell types. These mediums are serum free

Future Fields are using a biomanufacturing platform, the EntoEngine™, the first synthetic biology system in the world to use fruit flies like Drosophilia melanogaster for recombinant protein production to develop serum free growth factors for cell cultures for sustainable researchers. Their mission is to implement a sustainable biomanufacturing platform supporting the green initiatives.

Lectin Binding Assays

Vector laboratories offers over 125 lectin binding assays that are produced from plants and fungi. These lectins are highly purified, subjected to intensive quality control to ensure best availability of lectins for research purpose. These lectins can be used in IHC, ICC, IF, Blotting and ELISA applications. Lectin products are independently assessed by the National Centre for Functional Glycomics (NCFG) using the NCFG glycan arrays and the Consortium for Functional Glycomics (CFG) glycan arrays, providing researchers with detailed glycan-binding specificity and detailed data of complex carbohydrate structures and lot specific data for better reproducibility.

Industry thoughts on commercially available animal free antibodies, Pivotal Scientific, July 3, 2021; Industry thoughts on commercially available animal-free antibodies - Pivotal Scientific
Animal-free in vitro technologies, NC3Rs; Replacing animal-derived enzymes and cells for in vitro research | NC3Rs
Better antibodies without using animals, European Commission, 15 May 2020; Better antibodies without using animals (europa.eu)
The 3Rs, NC3Rs; The 3Rs | NC3Rs
Why drugs tested in Mice fail in Human Clinical Trials, Harvard University Article, February 11, 2020; Why Drugs Tested in Mice Fail in Human Clinical Trials - Science in the News (harvard.edu)

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Animal-Free Products

Recombinant Antibody and Proteins

Animal Free Diluent and Blocker Buffers

Animal Free Neural Cell Cultures Mediums

Lectin Binding Assays

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Animal-Free Products

Developments by Animal Welfare Organisations and Manufacturers

Key Benefits of Animal Free Products

Range of Animal Free Products by 2BScientific

Recombinant Antibody and Proteins

Animal Free Diluent and Blocker Buffers

Animal Free Neural Cell Cultures Mediums

Lectin Binding Assays

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