Expression of Single Chain Variable Fragment (scFv) Molecules in Plants: A Comprehensive Update
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REVIEW
Expression of Single Chain Variable Fragment (scFv) Molecules in Plants: A Comprehensive Update Padikara Kutty Satheeshkumar1
© Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Single chain variable fragments (scFvs) are generated by joining together the variable heavy and light chain of a monoclonal antibody (mAb) via a peptide linker. They offer some advantages over the parental mAb such as low molecular weight, heterologous production, multimeric form, and multivalency. The scFvs were produced against more than 50 antigens till date using 10 different plant species as the expression system. There were considerable improvements in the expression and purification strategies of scFv in the last 24 years. With the growing demand of scFv in therapeutic and diagnostic fields, its biosynthesis needs to be increased. The easiness in development, maintenance, and multiplication of transgenic plants make them an attractive expression platform for scFv production. The review intends to provide comprehensive information about the use of plant expression system to produce scFv. The developments, advantages, pitfalls, and possible prospects of improvement for the exploitation of plants in the industrial level are discussed. Keywords Recombinant protein · Targeted expression · Glycoengineering · scFv
Introduction Antibodies are used extensively as medicines, diagnostic molecules, environment clean-up agents, biosensors, and as a bait in the purification process in various commercial industries [1–3]. Due to the large size, complex nature, and extensive post-translational modifications, monoclonal antibodies (mAbs) with therapeutic applications are produced mostly using the mammalian expression systems. The cost factor involved in maintaining the manufacturing unit according to the GMP (Good Manufacturing Practise) regulations makes the commercial production of mAb an exorbitant process. A single chain variable fragment (scFv) is the smallest fragment of an antibody with the same antigen-binding specificity [4, 5]. Multimeric scFv comprising of more than one pair of heavy and light chains can be generated by changing the length of peptide linker. Multimerization can also be made by using specific peptides which have the natural ability to induce it [6, 7]. Multivalent scFv showing affinity to more than one protein at the same time * Padikara Kutty Satheeshkumar [email protected] 1
Centre of Advanced Study in Botany, Institute of Science, Banaras Hindu University, Varanasi, UP 221005, India
can be generated through genetic engineering [8–11]. The multivalent scFv binds to multiple antigens at the same time and can improve the diagnostic accuracy in any antibodydependent procedure. The concept and practice of using plants as recombinant protein expression platforms have made significant progress in the last two decades. There are many antibodies expressed successfully in plants [12–15]. The differences in the posttranslational modifications between mammalian cells and the plant ce
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