Gene cloning and characterization of a novel recombinant 40-kDa heat shock protein from Mesobacillus persicus B48

Fathinejad, Fatemeh; Ghafouri, Hossein; Barzegari, Ebrahim; Sarikhan, Sajjad; Alizadeh, Ali; Howard, Newton

doi:10.1007/s11274-023-03693-2

Cited by 1 publication

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“…Comprehensive research on fed-batch techniques in CHO cells has shown that the use of micronutrients can induce cell apoptosis, which is detrimental to recombinant protein production at high cell densities [ 19 ]. Some studies have indicated that overexpressing HSPs, such as HSP27, HSP40 and HSP70, can increase protein expression and cell viability [ 20 – 22 ]. In one study, CHO cells engineered to overexpress HSPs showed increased interferon-gamma production, although cell viability decreased with longer culture durations [ 23 ].…”

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confidence: 99%

Induction of heat shock protein expression in SP2/0 transgenic cells and its effect on the production of monoclonal antibodies

Jaffaraghaei,

Ghafouri,

Vaziri

et al. 2024

PLoS ONE

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The objective of the current investigation was to evaluate the induction of heat shock proteins (HSPs) in SP2/0 transgenic cells and the effect of these proteins on the production of monoclonal antibodies (mAbs). The SP2/0 cell line expressing the PSG-026 antibody, a biosimilar candidate of golimumab, the culture parameters, and the target protein expression were not justified for industrial production and were used for the experiments. Paracetamol and heat shock were used as chemical and physical inducers of HSPs, respectively. The results showed that paracetamol and heat shock increased the expression of HSP70 and HSP27 at the mRNA and protein levels. The expression of HSPs was greater in paracetamol-treated cells than in heat shock-treated cells. Paracetamol treatment at concentrations above 0.5 mM significantly reduced cell viability and mAb expression. However, treatment with 0.25 mM paracetamol results in delayed cell death and increased mAb production. Heat shock treatment at 45°C for 30 minutes after enhanced mAb expression was applied after pre-treatment with paracetamol. In bioreactor cultures, pretreatment of cells with paracetamol improved cell viability and shortened the lag phase, resulting in increased cell density. The production of mAbs in paracetamol-treated cultures was markedly greater than that in the control. Analysis of protein quality and charge variants revealed no significant differences between paracetamol-treated and control cultures, indicating that the induction of HSPs did not affect protein aggregation or charge variants. These findings suggest that inducing and manipulating HSP expression can be a valuable strategy for improving recombinant protein production in biopharmaceutical processes.

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