Laszlo

Hi I'm new here.

My name is Laszlo Groh, I'm an undergrad in the Netherlands currently on an internship working in Paediatric Infectious Diseases research laboratory.

I'm developing a bioassay using HEK-Blue cells from InvivoGen to detect soluble MDP in nasopharyngeal aspirate samples (NPA) collected from childeren (not going into anymore details ).

MDP is composed of N-acetylmuramic acid linked by its lactic acid moiety to the N-terminus of an L-alanine D-isoglutamine dipeptide (according to N. Inohara et al.)

There appears to be some contamination in the NPA samples that we are testing, and so we'd like to pre-treat our samples with heat or irradiation (UV or otherwise) inactivation. Our main concern is that we would like to choose a methodology of inactivation that has the least effect on MDP's conformational state (which is essential for our assay) yet maximum protein denaturation/deactivation.

So my question is, how much can I beat up MDP before is becomes biologically inactive? Heat vs. UV/Irradiation

Cheers,
Laszlo Groh