This is not the document you are looking for? Use the search form below to find more!

Report home > Science

Measurement of Electrostatic Interactions of Proteins Using Multi-Channel Surface Plasmon Resonance

4.65 (20 votes)
Document Description
Fee, C.J., Billikanti, J.M. and Damodaran, V. B. (2008) Measurement of Electrostatic Interactions of PEGylated Proteins Using a Novel Multi-Channel Surface Plasmon Resonance Technique. Recovery of Biological Products XIII, Qubec City, Qubec: 22-26 Jun 2008. (Conference Contribution %u2013 Poster presentation)
File Details
  • Added: March, 18th 2010
  • Reads: 1554
  • Downloads: 45
  • File size: 517.85kb
  • Pages: 1
  • Tags: spr, protein, surface interactions
  • content preview
Submitter
  • Name: DVB
Embed Code:

Add New Comment




Showing 17 comments

by Ann on March 20th, 2010 at 06:22 am
Comprehensive, but very informative
by Jane McCurdy on April 14th, 2010 at 11:23 pm
Interesting work
by Nathan Cornell on April 14th, 2010 at 11:29 pm
Awesome, I doubt whether these chips can be re-usable?
by K. G. R. Nair on April 15th, 2010 at 11:41 pm
Results look great. But whether these results were validated with other real-time biosensing devices like QCM?
by Andrea on June 25th, 2010 at 10:47 am
Nice one
by Jonathan on June 30th, 2010 at 07:09 pm
Why cation exchange only? Did you tried for anion exchange as well?
by Barb on July 06th, 2010 at 07:45 pm
Nice job
by Mike on August 24th, 2010 at 10:56 pm
It is really good to upload like this
by Kate on September 25th, 2010 at 10:21 am
Good
by Maria on October 01st, 2010 at 12:54 pm
Is there any correlation or possible to scaleup using ion exchange chromatography?
by Martha on October 09th, 2010 at 04:51 pm
I liked this
by Jennifer Christy on October 13th, 2010 at 10:25 am
Novel and interesting
by Catherine on October 19th, 2010 at 04:29 pm
Looks fine!!!
by Kimberly on October 21st, 2010 at 03:04 pm
Keep going, good!!!!!
by Chu on October 23rd, 2010 at 07:22 pm
Really awesome!!
by Bishop on October 24th, 2010 at 08:18 pm
Just average!!!
by Michell on November 11th, 2010 at 04:28 pm
Hi there, no error bars in any of your figures?!
Related Documents

Surface Plasmon Resonance Scattering and Absorption of anti-EGFR ...

by: sylwester, 7 pages

Gold nanoparticles with unique optical properties maybe useful as biosensors in living whole cells. Using a simple and inexpensive technique, we recorded surface plasmon resonance (SPR) scattering ...

Developing Effective Multi-Channel Marketing Strategies

by: rika, 12 pages

Although multi-channel marketing is the current trend, it is not an entirely new concept. Multi-channel has roots in the age-old “media mix” idea , which essentially said that buyers we ...

Infosys-Driving Multi-channel Commerce

by: manisha229249, 8 pages

Online shopping is being refined. Today, digital consumers rely heavily on multiple channels to research products, seek out information, and purchase goods and services. Read Infosys "Driving ...

Multi-channel Experience To Enhance Customer Satisfaction

by: 247inc, 1 pages

Giving customers the ability to meet their goals in their preferred channel, whether self-service or agent-assisted, improves the customer experience. At every touch point, an experience occurs and ...

Using Multi Layering To Design Interactions For Teaching

by: chang, 7 pages

Using Multi Layering to Design Interactions for teaching and learning in a Virtual Environment Platform – www.mycosm.com Invite only ...

Effects of Terms of Trade Gains and Tariff Changes on the Measurement of U.S. Productivity Growth

by: shinta, 51 pages

Since 1995, growth in productivity in the United States has accelerated dramatically, due in large part to the information technology sector. In this paper we argue that part of the apparent ...

Refractometer is a laboratory or field device for the measurement of an index of refraction

by: stephenwin, 0 pages

Atago USA is a company that sells refractometers, A refractometer is a laboratory or field device for the measurement of an index of refraction (refractometry). The index of refraction is calculated ...

The Measurement of Everyday Cognition (ECog): Scale Development and Psychometric Properties

by: shinta, 14 pages

This article describes the development and validation of an instrument to assess cognitively mediated functional abilities in older adults, Everyday Cognition (ECog). The ECog is an ...

Are maximizers really unhappy? The measurement of maximizing tendency

by: shinta, 7 pages

Recent research suggesting that people who maximize are less happy than those who satisfice has received considerable fanfare. The current study investigates whether this conclusion reflects ...

Economic Value Added and the Measurement of Financial Performance

by: bizmana, 3 pages

Shows how to calculate the appropriate discount rate for capital budgeting and other valuation problems. We now consider the measurement of fi nancial performance. We introduce the concept of ...

Content Preview
Measurement of Electrostatic Interactions of
Proteins Using Multi-Channel
Surface Plasmon Resonance

Conan J. Fee, Rayleen Fredericks-Short, Jaganmohan Billakanti, and Vinod Babu Damodaran
Department of Chemical & Process Engineering and the Biomolecular Interaction Centre,
University of Canterbury, Christchurch, New Zealand
Introduction and Objective
Cytochrome C 2B4Z
The charge-pH behaviour of proteins is of interest for
30.00
ion exchange processes. In this poster, we describe a
Native
new technique that utilises a multi-channel surface
20.00
e
1 PEG
2 PEG
plasmon resonance (SPR) instrument to measure
a
r
g
10.00
3 PEG
electrostatic interactions of cationic proteins in solu-
t
Ch

0.00
4 PEG
Ne
0
2
4
6
8
10
12
14
tion under a range of conditions.
-10.00
5 PEG
6 PEG
-20.00
Multi-Channel SPR
pH
Figure 1 shows the matrix arrangement of the ProteOn
Figure 3. Prediction of net charge-pH behaviour of na-
XPR36 SPR system (Bio-Rad Laboratories, Hercules,
tive and PEGylated Cytochrome C, calculated from PDB
CA), with 6 ligand channels and 6 analyte channels
sequence [1] using PropKa [2].
flowing at right angles. Cationic and anionic channels
can be prepared and proteins flowed across each si-
multaneously to measure interactions.
Relating SPR Interaction to Charge
From protein molecular weight and predictions of net
charge, Z, (Figure 3) at a given pH, the initial responses of
the cation exchange SPR to the proteins (Figure 4) were
related to the charge concentration in solution (Figure 5).
Cytochrome C

e

140
p
lo

120
S
m

100
r
a
r
g

]
o
s
80
Figure 1. Multi-Channel SPR. Individual channels can
s
n
e

[
RU/

60
be used for cation exchange, anion exchange and con-
l S
ia

40
it
In

trol surfaces.
20
R
P
S

0

0
0.2
0.4
0.6
0.8
1
Concentration [μM]
Chip Surface Preparation
Figure 4. Initial slope of SPR response for various Cyto-
A 1000 Da MW α-amino, ω-carboxyl-PEG was im-
chrome C concentrations on the cation exchange SPR chip.
mobilised on the surface of a GLC SPR chip (Bio-
Rad Laboratories) by amine coupling at pH 8.5 for 24
Cation Exchange SPR Response
hours. After blocking for 24 hours with ethanola-
Initial slopes of the SPR response were related to the charge
mine, taurine was amine coupled to the tethered car-
concentration in solution for Cytochrome C, Lysozyme and
boxy-PEG for 24 hours, floowed by further blocking.
Bovine Lactoferrin at three pH values (Figure 5). A
Taurine contains a sulfonic acid group that forms a
straight-line relationship exists, suggesting that the method
strong cation exchanger on the chip surface. Figure 2
may be useful for determining the net charge of unknown or
shows the SPR response to BSA (no interaction) and
modified (e.g. PEGylated) proteins.
Cytochrome C at pH 8.5 with (a) a native chip sur-

face and (b) the taurine surface. The native chip sur-
140
face contains carboxyl groups, so shows cation ex-
Lactoferrin pH 8.5
change activity.
Lactoferrin pH 7.5
120
Lactoferrin pH 6.5
]
/s

Cytochrome C pH 8.5
BSA
U
Cytochrome C pH 7.5
R
(a)
Cyt C 10,000 ng/mL
[ 100
Cytochrome C pH 6.5
Cyt C 5,000 ng/mL
e
Cyt C 2,500 ng/mL
s
Lysozyme pH 8.5
Cyt C 1,250 ng/mL
n
o

Running Buffer
p
Lysozyme pH 7.5
s
e

80
Lysozyme pH 6.5
R
R2 = 0.9525
R
P
l S

60
ia
it
f
In
o
e

40
p
lo
S

20
BSA
Cyt C 10,000 ng/mL
0
Cyt C 5,000 ng/mL
Cyt C 2,500 ng/mL
(b)
Cyt C 1,250 ng/mL
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
Running Buffer
Charge Concentration [Z/mL]
Figure 5. Cation exchange interaction as a function of the
sum of net charge and concentration.
References:
[1] PDB ID:2B4Z Mirkin, N., Jaconcic, J., Stojanoff, V., Moreno, A. (2008) High resolution X-ray crystallographic structure of bovine
Figure 2. Cation exchange SPR interactions (a) native
heart cytochrome c and its application to the design of an electron transfer biosensor. Proteins 70: 83-92 .
[2] Hui Li, Andrew D. Robertson, and Jan H. Jensen (2005). Very Fast Empirical Prediction and Interpretation of Protein pKa Values. Proteins,
chip (b) taurine-modified chip.
61, 704-721.


Download
Measurement of Electrostatic Interactions of Proteins Using Multi-Channel Surface Plasmon Resonance

 

 

Your download will begin in a moment.
If it doesn't, click here to try again.

Share Measurement of Electrostatic Interactions of Proteins Using Multi-Channel Surface Plasmon Resonance to:

Insert your wordpress URL:

example:

http://myblog.wordpress.com/
or
http://myblog.com/

Share Measurement of Electrostatic Interactions of Proteins Using Multi-Channel Surface Plasmon Resonance as:

From:

To:

Share Measurement of Electrostatic Interactions of Proteins Using Multi-Channel Surface Plasmon Resonance.

Enter two words as shown below. If you cannot read the words, click the refresh icon.

loading

Share Measurement of Electrostatic Interactions of Proteins Using Multi-Channel Surface Plasmon Resonance as:

Copy html code above and paste to your web page.

loading