Development of stable, serum-free suspension HEK293T cells for continuous Chikungunya VLPs production

Chikungunya Virus (CHIKV) infections occur in over 100 countries. Virus-like particles (VLPs), which replicate the structure of native virus without containing any infectious genome, hold promise as prophylactic vaccines. However, production of CHIK-VLPs using traditional HEK293T adherent cell culture, which relies on use of fetal bovine serum (FBS), faces scalability limitations. To address this challenge, we established a stable HEK293T cell line capable of producing CHIK-VLPs and further adapted it to suspension culture under serum-free conditions, improving scalability while reducing the risk of contamination associated with FBS. Further, a temperature shift during the process from 37 °C to 30 °C was incorporated to enhance the product yield. To confirm VLPs expression, various analytical techniques were employed, including electron microscopy, which assessed the morphology of VLPs produced by both stable adherent and suspension-adapted HEK293T cells. A 5-fold enhancement was achieved in suspension-adapted culture using a temperature shift, quantified by absorbance-based method. The established HEK293T cell line was capable of efficient CHIK-VLPs secretion provides a versatile and scalable method for VLPs production, it is also independent of antibiotics and FBS. This process represents a valuable contribution to the biopharmaceutical industry, offering potential for large-scale VLPs-based vaccine development.