Gene transformation and chain accumulation of the human monoclonal antibody adalimumab in Chlamydomonas reinhardtii.

• Grant number: 22/15451-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: February 01, 2023
• End date: December 31, 2023
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Lívia Seno Ferreira Camargo
• Grantee: Ednilson Donisete de França Junior
• Host Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil
• Associated research grant: 16/12992-6 - Chlamydomonas reinhardtii as a production platform to obtain biopharmaceuticals of national interest, AP.JP

Abstract

Genetic editing techniques made it possible for several proteins to be expressed heterologously. Currently, many proteins/peptides have already been produced recombinantly, using vectors such as: bacteria, yeast, mammalian cells, insect and plant cells. Complex recombinant biological medicines only benefit from a portion of these possible platforms, with the predominance of the use of mammalian cells in the current production scenario. This limitation is highly associated with the structural complexity and compatibility required in a protein for therapeutic purposes. Although animal cells overcome these problems, they can have a high maintenance/production cost with a low expression efficiency. Thus, it is important to study alternative platforms that are able to compete efficiently producing complex proteins. The microalgae Chlamydomonas reinhardtii has been extensively studied in the area of genetic engineering, presenting characteristics that impact production cost, level of recombinant expression and scaling. Among these characteristics we have photosynthetic capacity, accelerated growth, low cost of cultivation, high plastoma ploidy, existence of high expression endogenous regulatory elements, ability to carry out post-translational modifications and others. In this project, we aim at the genetic transformation of the genome of the C. reinhardtii chloroplast for the expression of one or two expression vectors at the same time, containing chains or the entire sequence of the adalimumab antibody, allowing the comparison of the expression of these chains and of the assembled antibody, under regulation of the psbA or psbD promoters. Adalimumab, a complex human monoclonal antibody that inhibits tumor necrosis factor, is included in the 2022 National List of Essential Medicines (Rename), an instrument used by the Unified Health System (SUS) to control pathologies in Brazil. Genetic modification and recombinant chain expression data will be obtained by applying molecular biology techniques, such as: PCR, enzymatic digestion, cloning techniques, SDS-PAGE and western blotting.