POTENTIAL BENCH TOP TECHNIQUE FOR MEASURING PROTEIN STABILITY IN SACCHARIDE-BASED GLASSES.

Jerainne Johnson, Jitendra Sharma and Marcus Cicerone

Proteins are stored dry in saccharide-based glasses for numerous biopharmaceutical and other biotechnological purposes. In spite of slow primary relaxation that occurs, aggregation and chemical degradation of proteins encapsulated in these glasses are persistent problems. Techniques commonly used to predict protein stability, thus far, have had mixed results. Recent neutron scattering experiments of these saccharide based glasses have shown that there is a correlation between protein stability in these glasses and fast local dynamics they display. Recent MD simulations and corresponding experiments have also shown a distinct correlation between protein stability and H-bonding dynamics occurring in the glass. Here we will show, with the use of the H-bond probe resorufin, steady state and time resolved fluorescence spectroscopy, that the above relationship holds true and can be used to establish a simple bench top methodology for evaluating host glass matrix materials that will enhance protein stability by suppressing such degradation processes as protein aggregation.

Mentor name:               Marcus Cicerone

Division:                       Polymers

Laboratory:                  MSEL

Room and Building

address:                        106 B/224

Mail Stop:                    8543

Telephone #:                 301-975-8595

FAX #:             301-975-4977

email:                            jerinne.johnson@nist.gov

Sigma Xi member:        No

Category:                     Biotechnology, Chemistry