1
2
The topic of today’s webinar is to introduce the MEA 2 Personal Purification
System. This is used exclusivley with PhyTi...
3
Analytical tools require small amounts of sample. However, the samples must
be pure. PhyNexus has identified a major bottl...
Protein purification is not automated for any significant throughput. High
throughput systems lack performance.
5
6
Utilize high throughput robotics by incorporating chemistry.
7
The PhyTip column is the core technology.
Flexible: tip volume, resin bed volume.
The back-and-forth flow is cornerstone...
PhyNexus is committed to installing solutions. The variety of resin bed sizes
gives the ability to fine tune yield and fin...
Equilibrium binding for scale-up
9
Efficient wash. Low dead volume, large dilution of interstitial space.
10
Placement of resin and screens means low dead volume and high
concentration elution. Resin is mostly water, gentle for pro...
13
Flexible technology for many applications. High performance due to column
design.
13
Compatible robotic systems.
14
System dedicated to processing PhyTip columns. Easy to use, does not require
specialized FTE. Versatile. Small footprint. ...
1616
The standard MEA 2 is equipped with a 1200 and 200 µL 12-channel head,
software and computer. The MEA is equipped with a c...
The MEA deck accommodates tips, reservoirs and plates.
18
19
20
IEX is possible by using step gradient elutions. Find conditions for large
isotherms. The protein is either on the resin o...
Conditions screening. In one experiment, study 288 conditions. Only possible
way of feasibly performing DOE studies.
22
23
Method development. Chromatographic separations.
Scale-up. Predictable results because of equilibrium binding. Screen conditions
and use results to design scale-up purific...
Merck was able to reproduce their two production scale columns using an 80 uL
and a 40 uL PhyTip column.
In this way, hund...
1000-fold smaller column. Get scaleable results and comparable purity.
26
Study many buffers at once. Screen for both conditions of high yield and high
activity. These experiments are not possible...
28
The MEA uses a 12-channel system. It is very straight forward compared to
FPLC systems.
29
Scale-able because of linear relationship between recovery and resin bed
volume. This is because of equilibrium binding.
30
Utilize specialized labware to process large volumes. The MEA 2 is capable of
processing 80 mL samples. In practice, most ...
High volume purification is comparable to gravity with the advantage of
automation and parallel processing. Different affi...
33
Obtaining crystals that diffract is not trivial. The protein must be soluble,
homogenous and high concentration. Proteins ...
Same approach was taken at Celgene.
35
36
PhyTip columns have the advantage of high performance. This is needed to
utilize the most effective downstream analysis...
High performance is demonstrated by comparison with spin columns. The
capability of loading a small resin bed more and elu...
Reproducibility between tip to tip. CV values of 10 or less.
39
40
41
42
Gel filtration chromatography through gravity flow. Fractionation shown here is
an example of capabilities.
43
Buffer exchange for dye labeling and excess dye removal. CE-LIF application
benefits from smaller volumes and higher th...
Sample prep such as glycan analysis is fully automated.
44
Using PhyTip columns packed with normal phase resin, thederivatized glycans
are enriched.
45
An example of glycan analysis using the HPCL.
46
47
48
Current HTP plasmid approaches require sample supernatent be transferred ro
plates or vacuums.
49
Using PhyTip columns, the crude sample san be processed without clearing the
supernatant, first.
DNA quality is copatible with sequencing, agarose gel analysis and transfection.
50
DNA used for expression screening.
51
52
53
PhyTip columns are used at every stage of the drug development process.
54
55
56
57
58
59
6060
Upcoming SlideShare
Loading in …5
×

The MEA 2 Personal Purification System for enhanced automated protein purification and sample preparation

484 views

Published on

To meet industry needs and customer requests, PhyNexus has developed the next generation MEA Personal Purification System. It is designed to process PhyTip columns for a range of automated chromatography applications including:
- Cold room operation for the purification of membrane and structural proteins.
- Fully automated plasmid and genomic DNA purification.
- Optimisation of process development e.g. for ion exchange purification protocols.
- Preparative-scale protein purification i.e. processing up to 12 samples in parallel of volumes as large as 60 mL.

Published in: Technology
0 Comments
0 Likes
Statistics
Notes
  • Be the first to comment

  • Be the first to like this

No Downloads
Views
Total views
484
On SlideShare
0
From Embeds
0
Number of Embeds
2
Actions
Shares
0
Downloads
0
Comments
0
Likes
0
Embeds 0
No embeds

No notes for slide

The MEA 2 Personal Purification System for enhanced automated protein purification and sample preparation

  1. 1. 1
  2. 2. 2 The topic of today’s webinar is to introduce the MEA 2 Personal Purification System. This is used exclusivley with PhyTip columns for complete purification and sample prep solutions.
  3. 3. 3
  4. 4. Analytical tools require small amounts of sample. However, the samples must be pure. PhyNexus has identified a major bottleneck in sample preparation. The solution was to miniaturize, process in parallel and automate. 4
  5. 5. Protein purification is not automated for any significant throughput. High throughput systems lack performance. 5
  6. 6. 6 Utilize high throughput robotics by incorporating chemistry.
  7. 7. 7 The PhyTip column is the core technology. Flexible: tip volume, resin bed volume. The back-and-forth flow is cornerstone of technology. Unique process for increased throughput. Placement of resin and low dead volume screens for high performance capture, efficient wash and high concentration elution.
  8. 8. PhyNexus is committed to installing solutions. The variety of resin bed sizes gives the ability to fine tune yield and final concentration.This variety of resin bed volumes is for specific requirements. PhyNexus installs solutions based on downstream needs and performance requirements. 8
  9. 9. Equilibrium binding for scale-up 9
  10. 10. Efficient wash. Low dead volume, large dilution of interstitial space. 10
  11. 11. Placement of resin and screens means low dead volume and high concentration elution. Resin is mostly water, gentle for proteins and required to maintain activity. 11
  12. 12. 13 Flexible technology for many applications. High performance due to column design. 13
  13. 13. Compatible robotic systems. 14
  14. 14. System dedicated to processing PhyTip columns. Easy to use, does not require specialized FTE. Versatile. Small footprint. Low maintenance. Just as reliable and robust as large systems, but less intimidating. 15
  15. 15. 1616
  16. 16. The standard MEA 2 is equipped with a 1200 and 200 µL 12-channel head, software and computer. The MEA is equipped with a chiller and buffer reservoir holder. 17
  17. 17. The MEA deck accommodates tips, reservoirs and plates. 18
  18. 18. 19
  19. 19. 20
  20. 20. IEX is possible by using step gradient elutions. Find conditions for large isotherms. The protein is either on the resin or it’s off the resin. 21
  21. 21. Conditions screening. In one experiment, study 288 conditions. Only possible way of feasibly performing DOE studies. 22
  22. 22. 23 Method development. Chromatographic separations.
  23. 23. Scale-up. Predictable results because of equilibrium binding. Screen conditions and use results to design scale-up purification. 24
  24. 24. Merck was able to reproduce their two production scale columns using an 80 uL and a 40 uL PhyTip column. In this way, hundreds of fermentation conditions can be tested and the results are directly scale-able. These data are published. 25
  25. 25. 1000-fold smaller column. Get scaleable results and comparable purity. 26
  26. 26. Study many buffers at once. Screen for both conditions of high yield and high activity. These experiments are not possible if running one condition at a time. 27
  27. 27. 28
  28. 28. The MEA uses a 12-channel system. It is very straight forward compared to FPLC systems. 29
  29. 29. Scale-able because of linear relationship between recovery and resin bed volume. This is because of equilibrium binding. 30
  30. 30. Utilize specialized labware to process large volumes. The MEA 2 is capable of processing 80 mL samples. In practice, most high volume users process 10-60 mL. 31
  31. 31. High volume purification is comparable to gravity with the advantage of automation and parallel processing. Different affinity resins are displayed here. 32
  32. 32. 33
  33. 33. Obtaining crystals that diffract is not trivial. The protein must be soluble, homogenous and high concentration. Proteins are also engineered to crystallize be removing hydrophobic residues on the surface and fixing flexible regions. Screening was demonstrated, here, using the MEA. These data are from SGC. 34
  34. 34. Same approach was taken at Celgene. 35
  35. 35. 36 PhyTip columns have the advantage of high performance. This is needed to utilize the most effective downstream analysis such as SDS-PAGE. For construct screening, the data have to be relatively high concentration to be useful.
  36. 36. High performance is demonstrated by comparison with spin columns. The capability of loading a small resin bed more and eluting in a smaller volumes means higher concentration samples. Compared with spin columns, the 20 µL Phytip columns are able to bind as much protein as a 100 µL spin column. The real advantage is the low volume elution, which keeps the pure protein more concentrated. The 5 µL PhyTip column does not have enough capacity, but the tradeoff is higher pure protein concentrations complared to the 20 µL Phytip column. 38
  37. 37. Reproducibility between tip to tip. CV values of 10 or less. 39
  38. 38. 40
  39. 39. 41
  40. 40. 42 Gel filtration chromatography through gravity flow. Fractionation shown here is an example of capabilities.
  41. 41. 43 Buffer exchange for dye labeling and excess dye removal. CE-LIF application benefits from smaller volumes and higher throughput automation.
  42. 42. Sample prep such as glycan analysis is fully automated. 44
  43. 43. Using PhyTip columns packed with normal phase resin, thederivatized glycans are enriched. 45
  44. 44. An example of glycan analysis using the HPCL. 46
  45. 45. 47
  46. 46. 48 Current HTP plasmid approaches require sample supernatent be transferred ro plates or vacuums.
  47. 47. 49 Using PhyTip columns, the crude sample san be processed without clearing the supernatant, first.
  48. 48. DNA quality is copatible with sequencing, agarose gel analysis and transfection. 50
  49. 49. DNA used for expression screening. 51
  50. 50. 52
  51. 51. 53
  52. 52. PhyTip columns are used at every stage of the drug development process. 54
  53. 53. 55
  54. 54. 56
  55. 55. 57
  56. 56. 58
  57. 57. 59
  58. 58. 6060

×