Pathogen reduction in human plasma using an ultrashort pulsed laser

Pathogen reduction is a viable approach to ensure the continued safety of the blood supply against emerging pathogens. However, the currently licensed pathogen reduction techniques are ineffective against non-enveloped viruses such as hepatitis A virus, and they introduce chemicals with concerns of side effects which prevent their widespread use. In this report, we demonstrate the inactivation of both enveloped and non-enveloped viruses in human plasma using a novel chemical-free method, a visible ultrashort pulsed laser. We found that laser treatment resulted in 2-log, 1-log, and 3-log reductions in human immunodeficiency virus, hepatitis A virus, and murine cytomegalovirus in human plasma, respectively. Laser-treated plasma showed ≥70% retention for most coagulation factors tested. Furthermore, laser treatment did not alter the structure of a model coagulation factor, fibrinogen. Ultrashort pulsed lasers are a promising new method for chemical-free, broad-spectrum pathogen reduction in human plasma

2019 Scholars at Work Conference Program

Program for the 2019 Scholars at Work Conference at Minnesota State University, Mankato on March 29, 2019

Slow Internal Dynamics and Charge Expansion in the Disordered Protein CGRP: A Comparison with Amylin

AbstractWe provide the first direct experimental comparison, to our knowledge, between the internal dynamics of calcitonin-gene-related peptide (CGRP) and amylin (islet amyloid polypeptide, IAPP), two intrinsically disordered proteins of the calcitonin peptide family. Our end-to-end contact formation measurements reveal that in aqueous solution (i.e., in the absence of structure-inducing organic solvents) CGRP preferentially populates conformations with short end-to-end distances. However, the end-to-end distance of CGRP is larger than that of IAPP. We find that electrostatic interactions can account for such a difference. At variance with previous reports on the secondary structure of CGRP, we find that the end-to-end distance of the peptide increases with decreasing pH and salt concentration, due to Coulomb repulsion by charged residues. Interestingly, our data show that the reconfiguration dynamics of CGRP is significantly slower than that of human IAPP in water but not in denaturant, providing experimental evidence for roughness in the energy landscape, or internal friction, in these peptides. The data reported here provide both structural and dynamical information that can be used to validate results from molecular simulations of calcitonin family peptides in aqueous solution

SDS-PAGE and native PAGE analysis of control and USP laser-treated plasma proteins.

<p>(A) On the left is shown the SDS-PAGE of control and laser-treated plasma; on the right, for comparison, is shown the SDS-PAGE of laser-treated MCMV virus adapted from Tsen <i>et al</i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111673#pone.0111673-Tsen3" target="_blank">[13]</a> (reprinted with permission from the Society of Photo-Optical Instrumentation Engineers). Control (untreated) or USP laser-treated plasma were boiled in reducing buffer and separated on a 10% gel. Proteins were visualized by Coomassie blue stain. The solid arrow indicates the location of low mobility detergent-resistant aggregates; the dotted arrow indicates missing band(s) corresponding to aggregated proteins. (B) Native PAGE of control and laser-treated plasma. Control (untreated) or USP laser-treated plasma were separated on a 10% gel. Proteins were visualized by Coomassie blue stain. Arrows indicate location of low mobility detergent-resistant aggregates.</p

Dynamic light scattering measurements on the aggregation state of fibrinogen.

<p>Dynamic light scattering measurements on the aggregation state of fibrinogen.</p