Ensure you have chosen the right quality for your antibody discovery program:
Microsart® Mycoplasma Detection Kits
Upstream testing helps ensure downstream success.
As part of a successful antibody discovery program, you want to select healthy clones that express high levels of your target antigen to optimize your downstream screens. Mycoplasma contamination interferes with your cells’ metabolism, can affect the properties of your produced antibody, and may adversely affect your success long-term.
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Mycoplasma contamination is subtle and pervasive
Mycoplasma are small (0.15 µm-0.3 µm) bacteria with no cell wall and a flexible membrane. These characteristics of Mycoplasma mean that contamination is subtle. Their lack of a cell wall and flexible membranes make them difficult to detect using microscopes, and their small size allows them to proliferate to high concentration within mammalian cell culture without any noticeable turbidity or other change in cell culture appearance.
Estimates suggest that between 5% and 30% of the world’s cell lines may be contaminated with one or more of the several known species of Mycoplasma.
Mycoplasma contamination is difficult to avoid because the major sources of contamination are lab personnel, standard cell culture components (for instance, fetal and newborn bovine serum and swine-derived trypsin), and cross-contamination from other cell cultures.
Contamination of your culture may not be apparent until your cell line is lost or your antibody is tested.
What is happening in your cell culture that you can’t see?
Mycoplasma contamination affects cell culture systems in many ways. Some effects may include:
Change of gene expression/protein expression profiles
Inhibition of protein biosynthesis due to arginine starvation
Inhibition of cell proliferation due to starvation/competition for nutrients in media
Production of toxic metabolites or nutrient competition adversely affecting antibody production
Changes in cell membrane antigenicity
Induction of apoptosis
Penetration of host cells, leading to long-term contamination
Acidification of media due to glucose metabolism
Inhibition of cellular differentiation
Activation of immune cells
Inhibition of cell growth
DNA fragmentation due to nucleases produced by mycoplasma
Compromised production of viruses
Inhibition of cell metabolism
Reduction of transfection efficiency
The best treatment is prevention
Cross-contamination from other contaminated cell lines is one of the main sources of mycoplasma contamination. Elimination of mycoplasma contamination from a culture is almost impossible; mycoplasma is resistant to most antibiotics and can survive liquid nitrogen without cryopreservation techniques, contaminating other cells stored in liquid nitrogen.
The best way to prevent whole-scale mycoplasma contamination of your antibody-producing clones:
Routinely test your existing cell lines and discard any positive cultures
Routinely quarantine and test any new cultures from outside sources
Other qPCR-based Mycoplasma detection kits on the market don’t give you an answer in three hours that
is as efficient- minimize pipetting effort by using less tubes (test 10 samples with 12 tubes using Microsart Mycoplasma Detection Kits vs 24-28 tubes for other kits. Directly incorporate controls in to the sample mix
is as cost-effective
uses your existing real-time PCR intstrument
Microsart® Mycoplasma Detection Kits offer qPCR-based detection with an answer in three hours using
highly-specific TaqMan® probes to reduce the risk of false-positives
multiplex PCR for sample and inhibition control to reduce the risk of false-positives and save on handling and analysis
inactivated, noninfectious validation standards to reduce the risk of cross-contamination