Lancaster plant scientists have adopted a technique rarely used in plant science to ensure their research meets the highest ethical standards.

Elizabete Carmo-Silva, professor of crop physiology at Lancaster University, has spent decades working to improve the efficiency of Rubisco Activase, a key protein in plants’ photosynthetic process. Her research aims to ensure key food crops remain resilient in a rapidly warming planet, helping to bolster food security around the world.

Carmo-Silva has stayed up to date on innovative and evolving plant science research methods, but one aspect of her work always concerned her – the process of making antibodies to detect and quantify proteins in a plant. This is frequently done in plant sciences, for example, to characterize the abundance of photosynthetic proteins in crop plants. Traditionally the process of making antibodies for research involved the use of a small number of animals – typically rabbits – which would have to be killed in order for their blood products to be used.

“The ethics of our research are very important, we’ve used antibodies made in the traditional way by external suppliers in the past but were always feeling very uneasy about using animals to do so,” said Carmo-Silva, principal investigator for the Realizing Increased Photosynthetic Efficiency (RIPE) project. “A doctoral researcher on my team, Duncan Bloemers, suggested we use a novel recombinant system to produce our new antibodies, which bypasses the need for animals, so I said great! Let’s do that.”

In recent years, advances in technology using cell cultures have begun to enable recombinant antibody production. Recombinant antibodies are proteins containing segments of DNA from different organisms and produced by a cell culture – removing the need to use animals. Carmo-Silva first heard about the technology from The UK National Centre for the Replacement Refinement & Reduction of Animals in Research (NC3Rs).

Using the method adopted by Bloemers and Carmo-Silva, scientists can generate antibodies for their target proteins and make progress towards engineering more efficient photosynthesis without sacrificing any animals.

“This technology has existed for years but it has not been used much in plant research either because scientists don’t know that it’s an option or because it’s cost-prohibitive,” said Bloemers, former doctoral researcher in the Carmo-Silva lab and now a data analyst for LiNaEnergy. “We were looking to make various antibodies to detect four different cowpea isoforms, which, using the traditional method, would require animals. We adopted this technology to make our research more ethical, even at the higher cost.”

A new book chapter by Bloemers and Carmo-Silva outlines how researchers can apply this method to plant science work. The chapter, “Antibody Design for the Quantification of Photosynthetic Proteins and Their Isoforms” appears in Photosynthesis: Methods and Protocols, a new book sharing methods, protocols, and best practices with the photosynthesis research community.

This will be the third edition that Carmo-Silva has contributed to, and she is excited to share the production of antibodies for photosynthetic proteins using recombinant technology with others. The chapter includes how Bloemers and Carmo-Silva went about designing and developing the antibodies specific to the different isoforms of their protein of interest (Rubisco Activase), what was involved in the design, and how to validate that the proteins are being properly recognized by the antibodies.

“We are very thankful to those who have made it possible to use this more ethical strategy moving forward,” said Carmo-Silva. “Now that we have adopted it and use it in our lab, we hope others will use it as well.”

Carmo-Silva conducts Rubisco research as part of RIPE, an international research project that aims to increase global food production by developing food crops that turn the sun’s energy into food more efficiently.