Wednesday, September 22, 2010

Manufacturing Biologics with CHO Cells? What’s the Risk for Viral Contamination?

by Dr. Ray Nims

Chinese hamster ovary (CHO) cells are frequently used in the biopharmaceutical industry for the manufacture of biologics such as recombinant proteins, antibodies, peptibodies, and receptor ligands. One of the reasons that CHO cells are often used is that these cells have an extensive safety track record for biologics production. This is considered to be a well-characterized cell line, and as a result the safety testing required may be less rigorous in some respects (e.g., retroviral safety) than that required for other cell types. But how susceptible is the cell line to viral contamination?

There are a couple of ways of answering this question. One way is to examine, in an empirical fashion, the susceptibility of the cell type to productive infection by model exogenous viruses. This type of study has been conducted at least three times over the past decades by different investigators. Wiebe and coworkers (In: Advances in Animal Cell Biology and Technology for Bioprocesses. Great Britain, 1989; 68-71) examined over 45 viruses from 9 virus families for ability to infect CHO-K1 cells, using immunostaining and cytopathic effect to detect infection. Only 7 of the viruses (Table 1) were capable of infecting the cells. Poiley and coworkers (In Vitro Toxicol. 4: 1-12, 1991) followed with a similar study in which 9 viruses from 6 families were evaluated for ability to infect CHO-K1 cells as detected by cytopathic effect, hemadsorption, and hemagglutination. This study did not add any new viruses to the short list (Table 1). The most recent study was conducted by Berting et al. This study involved 14 viruses from 12 families. The viruses included a few known to have contaminated CHO cell-derived biologics in the past two decades, and therefore did add some new entities to the list in Table 1. Still, the list of viruses that are known to replicate in CHO cells is relatively short.

Chinese hamster cells possess an endogenous retrovirus which expresses its presence in the form of retroviral particles, however these particles have been consistently found to be non-infectious for cells from other animals, including human cells. This endogenous retrovirus therefore does not present a safety threat (Dinowitz et al. Dev. Biol. Stand. 76:210–207, 1992).

A second way of looking at the question of viral susceptibility of CHO cells is to examine the incidence and types of reported viral contaminations of manufacturing processes employing CHO cell substrates. This subject has been reviewed a number of times, most recently by Berting et al. The types of viral contaminants fill a fairly short list (Table 2). In most cases, the contaminations have been attributed to the use of a contaminated animal-derived raw material, such as bovine serum.

Sources: Rabenau et al.1993; Garnick 1996; Oehmig et al., 2003; Nims Dev. Biol. 123:153-164, 2006; Nims et al., 2008; Genzyme 2009..

Considering the frequency with which CHO cell substrates have been used in biologics production, this history of viral contamination is remarkably sparse. This is further testament to the overall safety of this particular cell substrate.

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