Proteomic Analysis for the Assessment of Different Lots of Fetal Bovine Serum as a Raw Material for Cell Culture. Part IV. Application of Proteomics to the Manufacture of Biological Drugs

Abstract

Fetal bovine serum (FBS) is the most widely used growth supplement for cell cultures, primarily because of its high levels of growth stimulatory factors and low levels of growth inhibitory factors. Maintaining successful and consistent cell fermentations can be difficult, as FBS is a complex natural product and may vary from lot to lot even from a single manufacturer. The quality and concentration of both bulk and specific proteins can affect cell growth. Quality control tools for FBS are relatively primitive and expensive given the complexity of the sample and the large amounts of FBS used. We undertook this study to examine whether proteomics could be used as a tool to analyze the variability of different fermentation processes. We hypothesized that inconsistent cell growth in fermentations could be due to the quality of FBS and that different lots of FBS had varying concentrations of proteins such as growth stimulatory factors, growth inhibitory factors, and/or other proteins that may correlate with cellular growth rate. To investigate whether this was the case, we grew three batches of adult retinal pigment epithelial cells (ARPE-19) using three different lots of fetal bovine serum (FBS-Ia, FBS-Ib, and FBS-II). We found that the growth rate of the culture was significantly and consistently higher in the FBS-II lot. To determine why the other lots promoted different growth properties, we used proteomic techniques to analyze the protein composition of the three lots. We then performed a time course study to monitor specific changes in individual proteins in the fermentation medium. The amount of several extracellular matrix and structural proteins, which are indicators of cell growth, increased over time. Alternatively, components supplied by the FBS addition, such as nutritional-related and cell-spreading-related proteins, decreased over time.

Physical Properties of Polypeptide Electrospun Nanofiber Cell Culture Scaffolds on a Wettable Substrate

“…For HEWL and BSA labeling, the protein concentration was 2 mg/mL. Carbonate buffer was diluted 1:100 in PBS after labeling, and the final protein concentration was adjusted to 2 mg/mL, the approximate concentration of BSA in cell culture medium; 10% fetal bovine serum (FBS) has a protein concentration of 3-4.5 mg/mL, and the concentration of BSA is 0.69-1.52 mg/mL [16]. For direct labeling of PLEY fibers, FITC will have reacted with free amino termini.…”

An improved method for growing neurons: Comparison with standard protocols

“…Several media (N2, DMEM/F12) supplemented with serum, most commonly from fetal bovine (FBS) or horse origin, are commonly used for neuronal cultures. However, the chemical composition of the animal serum is not fully defined and includes some factors not present in the brain; moreover, the production of commercial serum is prone to batch to batch variability 3,4 . For these reasons, the use of a chemically defined, serum-free medium is recommended for studies in which a complete control of the environment in the nutrient medium is desired 5 .…”

Isolation of all CD44 transcripts in human epidermis and regulation of their expression by various agents

“…Calcium has been shown to up-regulate differentiationand ribosome protein-related genes and down-regulate genes involved in metabolism, DNA repair, transcription, and translation in primary human keratinocytes [66]. FCS is known to contain many mitogenic and growth factors and it is used to stimulate growth of many cell types [67]. Plasmin has been shown to be involved in wound healing by promoting migration of epidermal keratinocytes coupled with enhanced phagocytic-killing and inhibition of proliferation, which may facilitate re-epithelialization following skin injury [68] and has been shown to play a role in the amplification of psoriasiform skin inflammation in mice [69].…”