Although significant amounts of time and effort are spent on culturing hybridoma cell lines, poor growth or even cell death still occurs occasionally. Many factors, including long-term storage, repeated freeze-thaw cycles, improper handling, and contamination can all lead to failure of hybridoma survival causing permanent loss of important mAbs. Hybridoma cell lines are also known to undergo gene mutations and rearrangements over time, leading to antibody heterogeneity and batch-to-batch variability – ie. hybridomas can contain additional heavy or light chain genes resulting in impaired affinity and specificity to the target antigen; hybridomas also may lose the chromosome containing the antibody genes required for antibody generation.
Next generation protein sequencing (NGPS) can provide the amino acid sequence of any antibody to help recover from the loss, death or mutation of hybridoma cell lines. Indefinite access to the amino acid sequence can mean a limitless supply of commonly used antibodies through recombinant expression, providing an insurance policy in case they are discontinued. NGPS can also validate every batch of antibodies produced by hybridomas, confirming batch-to-batch consistency.