Biophysical Society 59th Annual Meeting Program Guide

182

Biophysical Society 59

Annual Meeting, Baltimore, Maryland

Exhibitor Presentations

Exhibit Hall C, Baltimore Convention Center

Room A: Sunday, February 8

7:30

–9:00

FEI Company

FEI Cryo-TEM Workflow Solutions: A New Era for 3D Structural

Biology

A new frontier exists in unraveling interactive biological and biochemical

processes and pathways at the macromolecular level. Of critical importance

is the three-dimensional visualization of macromolecular structures and

molecular machines in their native functional state. Three techniques play a

major role in orchestrating this.

Nuclear magnetic resonance (NMR) has the capability to study specific

protein domains or fragments and their functional role in protein folding

and dynamics and in ligand binding whereas X-Ray crystallography (XRD)

allows visualizing high-resolution but more static 3D structures of apo and

liganded proteins, mainly in a monomeric or dimeric state after crystal-

lization. To unravel more physiologically relevant situations however, it is

essential to visualize multimeric complexes in their tertiary and quaternary

state and their interaction with other complexes. By performing typical

cryo-TEM applications like single particle analysis or tomography, this

can be achieved. In this so-called translational methodology, cryo-TEM

thus provides complementary information to NMR and XRD that can

be crucial for drug discovery, e.g. in terms of a better understanding of the

mechanism of action inferred from the EM structure of the physiologically

relevant complex. This will eventually contribute to answer real biologically

as well as medically relevant questions.

Latest developments in the cryo-TEM workflow have brought the 3 major

structural biology technologies closer together. Now, finally, a continuum

has been reached on all important aspects with regards to resolution and

macromolecular scales which allows for the full deployment of the combina-

tion of these technologies.

Here, we will illustrate the historical context of these technologies with

respect to one another and show how latest developments have reached the

critical requirements to fully unleash the power of structural biology in not

just answering fundamental questions, but actually contribute to curing

diseases and improving health. Also, we will discuss the future of struc-

tural biology based on the latest developments of the FEI workflow and its

components with a special focus on the advances in contrast enhancement

(phase plates) and (direct electron) detection.

Presenter

Chris Arthur , Applications Engineer, FEI Company

3:30

–5:00

Wyatt Technology Corporation

The Light Scattering Toolkit for Biophysical Characterization: Lab

Essentials for Enhancing Studies of Purification, Crystallization, For-

mulation, Conjugation, Conformation, and Interactions

Biophysical techniques based on static and dynamic light scattering address

many of the key analytical challenges associated with proteins, oligonucle-

otides, vesicles and other biomacromolecules. This workshop covers the

following topics:

1. Batch DLS—traditional cuvette-based dynamic light scattering (DLS)

is a fast, easymeans of estimatingmacromolecular and nanoparticle size

distributions to assess protein aggregation or the sizes of virus-like par-

ticles or drug delivery nanovehicles. In microwell-plate format, DLS is

a high-productivity tool useful for optimizing formulation or crystalli-

zation conditions with minimal sample consumption or manual labor.

2. SEC-MALS and SEC-DLS—coupling of multi-angle static light scat-

tering (MALS) and DLS detection to size-exclusion chromatography

to assess molar mass, size, conformation and conjugation, in solution,

independently of column calibration and non-ideal sample-column

interactions. In addition to readily assessing aggregation and fragmen-

tation in line with SEC purification, SEC-MALS analyzes protein

conjugates such as glycoproteins or membrane proteins bound to

surfactant micelles, determining protein oligomeric state and the

mass of glycans, polysaccharides or surfactant modifying the protein.

3. FFF-MALS and FFF-DLS—coupling of MALS and DLS to a field-

flow fractionation (FFF) device to achieve accurate characterization of

macromolecules and nanoparticles from 1-1000 nm, even when solu-

ble and insoluble components are both present in the solution. It does

not employ a stationary phase; FFF separates without shear and with

minimal surface interactions. FFF produces high-resolution size distri-

butions thanks to true hydrodynamic separation upstream of the light

scattering detectors. It also offers the benefits of post-separation down-

stream analysis by spectroscopy for additional information on samples.

4. CG-MALS—coupling MALS to a composition-gradient device

results in a uniquely powerful system for characterizing complex

biomolecular interactions, label-free and immobilization-free. Because

MALS measures molar masses it is one of the most useful techniques

for analyzing multi-domain, multi-protein interactions that go beyond

standard 1:1 interactions including systems exhibiting cooperativ-

ity and allostery. CG-MALS determines the affinity and absolute

molecular stoichiometry of self and/or heteroassociating systems from

pM to mM.

Presenter

Stephanie Cope, Applications Scientist, Wyatt Technology Corporation