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Transcript of 1-888-780-8535 © 2008 1-888-780-8535 © 2009 U.S. Patents #7,129,091, #6,409,832 and Patents...
![Page 1: 1-888-780-8535 © 2008 1-888-780-8535 © 2009 U.S. Patents #7,129,091, #6,409,832 and Patents Pending 1 The Microcapillary Protein.](https://reader035.fdocuments.in/reader035/viewer/2022062519/5697c0191a28abf838cce730/html5/thumbnails/1.jpg)
1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
1
The Microcapillary Protein Crystallization System
MPCSTM
This work supported in part by NIGMS NCRR co-sponsored PSI-2 Grant U54 GM074961
Cory GerdtsEnabling Technologies Workshop
3/4/09
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
MPCS Competitive Advantages:
• Low Volume (5-10 nL Protein / experiment)
• Diffraction-ReadyTM Crystals – translation / scale up not required
• Crystals easily extracted from Peel-ApartTM CrystalCards
• No Dead Volume – Every nL Gets Used
• In situ diffraction
• ~22 L performs a gradient of 96 conditions
2
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
Zheng B.; Roach L. S.; R. F. Ismagilov J. Am. Chem. Soc. 2003, 125: 11170-11171.
Plug-Based Protein Crystallization
Zheng, B.; Ismagilov, R.F. Angew. Chem. 2005, 117: 2576-2579.
H. Song, J. D. Tice, R. F. Ismagilov Angew. Chem.-Int. Edit. 2003, 42, 768.
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
CrystalCard™
20 nl Plugs200 m microcapillary
How Does it Work?
4
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
CrystalCard™
20 nl Plugs200 m microcapillary
5
How Does it Work?
![Page 6: 1-888-780-8535 © 2008 1-888-780-8535 © 2009 U.S. Patents #7,129,091, #6,409,832 and Patents Pending 1 The Microcapillary Protein.](https://reader035.fdocuments.in/reader035/viewer/2022062519/5697c0191a28abf838cce730/html5/thumbnails/6.jpg)
1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
![Page 7: 1-888-780-8535 © 2008 1-888-780-8535 © 2009 U.S. Patents #7,129,091, #6,409,832 and Patents Pending 1 The Microcapillary Protein.](https://reader035.fdocuments.in/reader035/viewer/2022062519/5697c0191a28abf838cce730/html5/thumbnails/7.jpg)
1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
Efficient Optimization of Crystal Growth by Fine Gradient Screening
pH 6 pH 7 pH 8
• Optimize crystal growth using fine control over chemical gradients for:
Protein, Buffers (pH), Salts, Ligands, Additives, Precipitants, etc.
20 nl Plugs
7
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
A Smooth Chemical Gradient with the MPCS
Single Crystal
Precipitate
Microcrystals
Single Crystals
Crystal Clusters
Gerdts et al. Acta Cryst. (2008). D64, 1116–1122.
8
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
Bups 27
SSGCID and the MPCS
Methionine-R-sulfoxide reductase of Burkholderia pseudomallei Bups.00033.a 1.7 Å PDB ID:
3CXK 4-24-08
Bups.00014.b.A1 2.9 Å PDB ID: 3EOO
Gerdts et al. Acta Cryst. (2008). D64, 1116–1122
Bups 69
Bups 35
Bups 75
Bups 33+ Met
Bups 14 Bups 33
9
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
![Page 11: 1-888-780-8535 © 2008 1-888-780-8535 © 2009 U.S. Patents #7,129,091, #6,409,832 and Patents Pending 1 The Microcapillary Protein.](https://reader035.fdocuments.in/reader035/viewer/2022062519/5697c0191a28abf838cce730/html5/thumbnails/11.jpg)
1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
Spacer Gas Bubble
Spacer
Protein
Li, L., et al. (2006). PNAS. 103, 19243.
• Pre-formed cartridges of crystallization reagents are combined with chemical gradients for efficient screening.
• ~22 L performs a gradient of 96 conditions (up to 3500 individual experiments)
Hybrid Method: Sparse Matrix + Fine Chemical Gradient Screening
11
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
No Dead Volume – Every nL is Used
• The Plug Maker aspirates protein into one of the tubes connected to a syringe that is back-filled with the carrier fluid (cf). Every nL of protein gets displaced into the CrystalCard.
cfSyringe filled with carrier fluid
12
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
detector
X-Rays
MPCS™ CrystalCard™
(Side view)
30°30°
In situ X-ray Diffraction Schematic
goniometer head
Beam Line Requirements:
In-Line Optics for Crystal Aligment
Stable and Accurate Motion Control
Small Intense Beam
Novel High Resolution Detectors
13
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
detector
goniometer head
X-Rays
MPCS™ CrystalCard™
(Side view)
30°30°
In situ X-ray Diffraction Schematic100 m1 mm
Beam Line Requirements:
In-Line Optics for Crystal Aligment
Stable and Accurate Motion Control
Small Intense Beam
Novel High Resolution Detectors
14
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
In situ X-ray Diffraction Screening and Data Collection
• Crystals grown in the
CrystalCard were
subjected - in situ – to X-
rays. Data collection at
the NE-CAT beamline
24ID-C at APS
• Data merged from 3
crystals to generate a 1.9
Å resolution electron
density map
• In situ Screening
performed at GM/CA 23B
• CrystalCard goniometer
holder being developed
Gerdts et al. Acta Cryst. (2008). D64, 1116–1122.
15
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
MPCS User Facility Visit
MPCS User Facility ~5 miles from APS at ANL
2501 Davey Road, Woodridge, IL 60517
www.emeraldbiosystems.com
Development of the NIGMS NCRR co-sponsored PSI-2 Grant U54 GM074961
16
MPCS-Beta Instrument
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
The Microcapillary Protein Crystallization System (MPCS™)
Plug Maker™ & CrystalCards™
PSI Appreciation
Discount: 30%
Contact us TODAY for a Quotation:[email protected]
In appreciation of the ATCG3D and PSI funding, we are introducing the MPCS with a 30% discount!
Discount offered for limited time only. Contact us for details.U.S. Patents #7,129,091, #6,409,832 and Patents Pending
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
• Touch-Screen User Interface
• Small Footprint (~18”x15”x14”)
• No Dead Volume
• Low Volume (5-10 nL Protein / Plug)
• ~22 L Performs Gradient of 96 Conditions
• Diffraction-Ready Crystals – “Translation” not Required
• Automated Crystallization Screening – Sparse Matrix and
Gradient Optimization
• Fully Integrated Experimental Setup (Hardware/Software)
The Plug Maker™
18
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
Automated Sampling
19
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
Loaded and Ready
20
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
MPCS Competitive Advantages:
• “I can’t get my crystals out”
21
MPCS: Other Microfluidic
Crystallization System(s):
• Peel-ApartTM CrystalCardsTM
• “Crystal hits don’t translate or
scale-up to vapor diffusion”
• Diffraction-ReadyTM crystals –
translation/scale-up unnecessary
• “Microchips are expensive”
(minimum ~$0.63 per drop)
• More than 3X cheaper
(~$0.19 per drop)
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending
www.emeraldbiosystems.com/MPCS
22
- Videos
- e-presentation (youtube)
- Slides
- User Facility
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1-888-780-8535 © 20081-888-780-8535 © 2009 www.emeraldbiosystems.com
U.S. Patents #7,129,091, #6,409,832 and Patents Pending