Written by María Gerpe, PhD
August 4, 2021
Antibodies are used in a variety of ways in academia and industry, from tools to therapeutics. Because antibodies are produced using live processes, which are naturally error-prone, validation is required from time to time (Lamanna et al. 2017). Furthermore, to develop biological therapeutics, the protein sequence must be confirmed as part of the regulatory process (Scott et al. 2014).
Why Do Antibodies Need Validation?
Antibody discovery systems are biological in nature, and are thus not immune to errors. Hybridomas, for instance, throughout time, may acquire aberrations that result in additional production of chains, or an antibody with a sequence that has deviated from the original sequence. Furthermore, sometimes hybridomas are not perfectly monoclonal by nature, and may in fact produce more than one antibody.
In the case of single B-cell sequencing, recombinant production is not exempt from eventually encountering problems down the road, as aberrations have been noted in the literature after several passages of cell lines. Phage, yeast and bacterial display carry their own issues, which other than potential DNA errors, include a lack of biologically relevant post-translational modifications (PTMs).
Antibodies are widely used in labs and therefore are quickly consumed; multiple lots from the “same” antibody will be used throughout a year at any given lab. Because lot-to-lot variability is a common issue, it is critical to validate antibodies generated in-house, and those bought commercially to ensure reproducibility of the research or treatment.