Metabolic profiling of insect cell lines: Unveiling cell line determinants behind system's productivity

Abstract

Baculovirus infection boosts the host biosynthetic activity towards the production of viral components and the recombinant protein of interest, hyper-productive phenotypes being the result of a successful adaptation of the cellular network to that scenario. Spodoptera frugiperda derived Sf9 and Trichoplusia ni derived High Five cell lines have a major track record for the production of recombinant proteins, with High Five cells presenting higher productivities. A metabolic profiling of the two insect cell lines was pursued to underpin specific cellular traits behind productive phenotypes. Multivariate analysis identified cell-line dependent metabolic signatures linked to productivity. Pathway analysis highlighted cellular pathways of paramount importance in supporting infection and protein production. Moreover, better producer phenotypes proved to be correlated with the capacity of cells to shift their metabolism in favor of energy-generating pathways to fuel biosynthesis, a scenario observed in the High Five cell line. Metabolomic profiling allowed us to identify metabolic pathways involved in infection and recombinant protein production, which can be selected as targets for further improvement of the system.

Entometabolomics: applications of modern analytical techniques to insect studies

“…The percentage is higher among post-2006 studies (12/33, 36.4%). One particular recent investigation (Monteiro et al, 2014) did not apply any form of statistical testing, instead using an arbitrary cut-off to conclude whether differences were present or absent. Although it is possible that this approach could accord with results of formal hypothesis testing, this is by no means certain, especially with low numbers of replicates (Quinn & Keough, 2002).…”

“…Phalaraksh et al (2008);34 Monteiro et al (2014);35 Buszewska-Forajta et al (2014b); 36 Buszewska-Forajta et al (2014a); 37 Wu et al (2012); 38 Lenz et al (2001); 39 Zhang et al (2007); 40Dossey et al (2006);41 Verberk et al (2013).…”

The role of host cell physiology in the productivity of the baculovirus‐insect cell system: Fluxome analysis of Trichoplusia ni and Spodoptera frugiperda cell lines

“…Sf9 cells have a very efficient oxidative metabolism, with the channeling of glucose toward pyruvate formation and further incorporation of this carbon source to the TCA cycle . Having this in consideration, the metabolomic profile of both cell lines was analyzed and correlated with their productivity phenotype (Monteiro et al, 2014). Regarding productivity, it is recognized that Sf9 cells are a better host for the production of infectious baculovirus vectors, while High Five cells are preferred for high level expression of recombinant proteins (Krammer et al, 2010;Taticek et al, 2001;Wilde et al, 2014).…”

“…Moreover, the low levels of ammonia produced by Sf9 cells are related with the presence of a detoxification and recycling mechanism driven by glutamate synthase (GOGAT) activity (Doverskog et al, 2000). Such analysis retrieved major differences on redox homeostasis and nucleotide biosynthesis, related with higher productivity of recombinant proteins by High Five cells (Monteiro et al, 2014). Having this in consideration, the metabolomic profile of both cell lines was analyzed and correlated with their productivity phenotype (Monteiro et al, 2014).…”

The components of shear stress affecting insect cells used with the baculovirus expression vector system

“…Depending on the genetic construct, the recombinant protein can accumulate in the cell or can be secreted into the medium. During the infection, the virus takes complete control of the host cell, increasing its catabolic metabolism and thus causing higher oxygen consumption [11,12]. The infected cells are arrested in the G2/M phase [13] and cytoskeleton is destabilized [14,15].…”