title: Production of functional active human growth factors in insects used as living biofactories.
abstract: Growth factors (GFs) are naturally signalling proteins, which bind to specific receptors on the cell surface. Numerous families of GFs have already been identified and remarkable progresses have been made in understanding the pathways that these proteins use to activate/regulate the complex signalling network involved in cell proliferation or wound healing processes. The bottleneck for a wider clinical and commercial application of these factors relay on their scalable cost-efficient production as bioactive molecules. The present work describes the capacity of Trichoplusia ni insect larvae used as living bioreactors in combination with the baculovirus vector expression system to produce three fully functional human GFs, the human epidermal growth factor (huEGF), the human fibroblast growth factor 2 (huFGF2) and the human keratinocyte growth factor 1 (huKGF1). The expression levels obtained per g of insect biomass were of 9.1, 2.6 and 3mg for huEGF, huFGF2 and huKGF1, respectively. Attempts to increase the productivity of the insect/baculovirus system we have used different modifications to optimize their production. Additionally, recombinant proteins were expressed fused to different tags to facilitate their purification. Interestingly, the expression of huKGF1 was significantly improved when expressed fused to the fragment crystallizable region (Fc) of the human antibody IgG. The insect-derived recombinant GFs were finally characterized in terms of biological activity in keratinocytes and fibroblasts. The present work opens the possibility of a cost-efficient and scalable production of these highly valuable molecules in a system that favours its wide use in therapeutic or cosmetic applications.
authors.affiliation: Alternative Gene Expression S.L. (ALGENEX), Centro empresarial, Parque Científico y Tecnológico de la Universidad Politécnica de Madrid, Campus de Montegancedo, Pozuelo de Alarcón, 28223 Madrid, Spain.
authors.forename: Benoit
authors.initials: B
authors.lastname: Dudognon
authors.affiliation: Alternative Gene Expression S.L. (ALGENEX), Centro empresarial, Parque Científico y Tecnológico de la Universidad Politécnica de Madrid, Campus de Montegancedo, Pozuelo de Alarcón, 28223 Madrid, Spain.
authors.forename: Lorena
authors.initials: L
authors.lastname: Romero-Santacreu
authors.affiliation: Alternative Gene Expression S.L. (ALGENEX), Centro empresarial, Parque Científico y Tecnológico de la Universidad Politécnica de Madrid, Campus de Montegancedo, Pozuelo de Alarcón, 28223 Madrid, Spain.
authors.forename: Silvia
authors.initials: S
authors.lastname: Gómez-Sebastián
authors.affiliation: Vivacell Biotechnology España S.L. Parque Científico Tecnológico Rabanales, 21, c/Cecilia Payne, Parcela ID 8.1, 14014 Córdoba, Spain.
authors.forename: Ana B
authors.initials: AB
authors.lastname: Hidalgo
authors.affiliation: Alternative Gene Expression S.L. (ALGENEX), Centro empresarial, Parque Científico y Tecnológico de la Universidad Politécnica de Madrid, Campus de Montegancedo, Pozuelo de Alarcón, 28223 Madrid, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: López-Vidal
authors.affiliation: Vivacell Biotechnology España S.L. Parque Científico Tecnológico Rabanales, 21, c/Cecilia Payne, Parcela ID 8.1, 14014 Córdoba, Spain.
authors.forename: María L
authors.initials: ML
authors.lastname: Bellido
authors.affiliation: Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Córdoba, Spain.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Muñoz
authors.affiliation: Department of Biotechnology, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Autovía A6, Km 7.5, 28040 Madrid, Spain. Electronic address: escriban@inia.es.
authors.forename: José M
authors.initials: JM
authors.lastname: Escribano
mesh_terms: D000818:Animals; D019149:Bioreactors; D004815:Epidermal Growth Factor / Q000096:biosynthesis* / Q000235:genetics; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis* / Q000235:genetics; D051523:Fibroblast Growth Factor 7 / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007814:Larva / Q000235:genetics / Q000378:metabolism; D009036:Moths / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D051523:Fibroblast Growth Factor 7; D004815:Epidermal Growth Factor
keywords: Baculovirus; Biofactory; Recombinant Growth Factors; Trichoplusia ni larvae