Foresight™
RoboColumn Units
200 and 600 µl
Instruction Manual
Catalog Numbers
732-4801 732-4816
732-4802 732-4817
732-4804 732-4819
732-4805 732-4820
732-4807 732-4822
732-4808 732-4823
732-4813 732-4825
732-4814 732-4826
Bio-Rad Technical Support
For help and technical advice, please contact the Bio-Rad
Technical Support department. In the United States, the Technical
Support department is open Monday–Friday, 5:00 AM to 5:00 PM,
Pacific Time.
Phone: 1-800-424-6723
Fax: 1-510-741-5802
Email: LSG_TechServ_US@bio-rad.com (for U.S. and
international customers)
Online technical support and worldwide contact information are
available at www.consult.bio-rad.com.
Legal Notices
RoboColumn is a trademark of Atoll GmbH.
Freedom EVO, Freedom EVOware, Te-Chrom, and Te-Stack are
trademarks of Tecan Group Ltd.
© 2012 by Bio-Rad Laboratories, Inc. All rights reserved.
Table of Contents
Bio-Rad Technical Support ii
Legal Notices ii
Section 1 Introduction 1
Section 2 Characteristics 2
Section 3 Advice on Handling 4
3.1 Equipment 4
3.2 Sample Preparation 6
3.3 Experiment Setup 6
3.4 Te-Chrom Wizard 7
Section 4 Protocol 8
4.1 Detailed Protocol 8
Section 5 Troubleshooting Guide 15
Section 6 Ordering Information 16
Section 1
Introduction
Foresight™ RoboColumn units are prepacked chromatography
microcolumns used for rapid process development. Their design
is fully compatible with robotic liquid handling workstations, such
as the Freedom EVO series from Tecan, and allows process
developers to perform high throughput (HT) parallel screening
and experimentation. Foresight RoboColumn units may be used
for qualitative media screening experiments and/or small scale
methods development. Through design of experiments (DoE)
utilizing Foresight RoboColumn units, an operational window can
be defined to aid in developing a quality by design (QbD) process.
The design of the Foresight column offers:
1. Convenience
The prepacked columns used in conjunction with a robotic
liquid handling device allow users to create automated
protocols for HT experimentation. The ability to define a design
space enables process scientists to develop robust purification
processes.
2. Reliability
Each column is individually packed under optimum
compression, ensuring consistent experimental results.
3. Efficiency
The small column design enables users to use precious
feed sample more efficiently while performing real
chromatographic separations in parallel to determine critical
quality parameters.
Foresight RoboColumn units are available in a variety of
chromatography modes including anion, cation, and mixed-mode.
See Section 6, Ordering Information, for the complete product line.
Technical Support: 1-800-424-6723 • www.bio-rad.com 1
Section 2
Characteristics
Foresight™ RoboColumn units are ready-to-use chromatography
microcolumns prepacked with Bio-Rad Laboratories process
chromatography resins. Each row contains eight columns
and either a single row or multiple rows may be used. For HT
screening, configuration into 96-well plate format is possible with
the use of an array plate.
Foresight RoboColumn units are available in two volumes:
200 and 600 µl.
Table 1. Characteristics of Foresight RoboColumn units.
Plate size 128.3 x 86.0 x 14.0 mm
Plate material Polyoxymethylene copolymer
Number of wells Polypropylene/polyethylene
Column inner diameter 5 mm
Column material Polypropylene
Chemical stability Stable in organic solvents. UNOsphere™ and
Avoid Halogenated organic solvents, hexane
Storage solution • UNOsphere Q, UNOsphere S, UNOsphere
Recommended
storage temperature
Working temperature 4–30°C
Maximum working
pressure
Flow velocity 16–1000 cm/h
Laser label Placed on every individual column body and
*
Specifications for all Bio-Rad chromatography media may be found by
visiting www.bio-rad.com/process and downloading their corresponding
instruction manual.
Nuvia™ resins are stable in commonly used
aqueous buffers pH 1–14. CHT™ resin is stable in
aqueous buffers of pH >60.5.*
Rapid S, Nuvia™ cPrime™: 20% ethanol
• CHT Type I, 40 µm and CHT Type II, 40 µm:
20% ethanol + 0.01M Na phosphate, pH 6.8
• Nuvia Q: 20% ethanol + 1M NaCl
• Nuvia S: 20 % ethanol + 0.1M NaCl
4–30°C
Up to 8.0 bar
2
containing:
• Product number
• Resin name
• Column volume in µl
2 Technical Support: 1-800-424-6723 • www.bio-rad.com
Table 2. List of Bio-Rad chromatography media.
Chromatography
Media
UNOsphere Q Strong Anion 4110109
UNOsphere S Strong Cation 4110109
UNOsphere
Rapid S
Nuvia Q Strong Anion 10018215
Nuvia S Strong Cation 10018215
Nuvia cPrime
CHT Type I, 40 µm
CHT Type II, 40 µm
Mode
Strong Cation 10010339
Mixed Mode
(Hydrophobic-Cationic)
Mixed Mode
(Metal Affinity-Cationic)
Mixed Mode
(Metal Affinity-Cationic)
Instruction Manual
Literature #
10023853
6086
6086
Technical Support: 1-800-424-6723 • www.bio-rad.com 3
Section 3
Advice on Handling
3.1 Equipment
Foresight™ RoboColumn units are designed for robotic handling.
Table 3 is a guide to the equipment required for robotic handling of
the units using Tecan’s Freedom EVO workstation.
Table 3. Recommended equipment for automated handling of
Foresight RoboColumn units.
Equipment Details Tips and tricks
RoboColumn units
96-well array plate
Te-Chrom
Module (A)
Arrange up to
96 individual
columns onto
the 96-well array
plate according
to application
requirements
Use the Te-Chrom
module to integrate
the Foresight
RoboColumn units
onto the robotic
worktable
• Height
adjustable
• Waste option
(B)
• Do not mix short
(200 µl) and long
(600 µl) columns
• Make sure the
columns are securely
seated into the
96-well array plate
before using them
• Use the Te-Chrom
module in
combination
with a Te-Stack
plate stacker for the
optimized handling
and storage of
collection plates
• Use the waste
container to remove
buffers from each
step
• Make sure that the
columns are an
appropriate distance
from the collection
plate before starting
the experiment
4 Technical Support: 1-800-424-6723 • www.bio-rad.com
Table 3. Recommended equipment for automated handling of
Foresight RoboColumn units, continued.
Equipment Details Tips and tricks
Te-Chrom
Shuttle
Te-Chrom
Wizard
Plate Reader Use the plate
Use the Te-Chrom
shuttle to move
a collection plate
under the columns
to fractionate the
flowthrough from
the column outlets
Use the
Te-Chrom Wizard
to configure the
chromatographic
process and to set
all process-relevant
parameters
(volume, flow rate,
etc.)
reader to evaluate
data such as UV
measurement
and protein
concentration
Use the Robotic
Manipulator Arm (RoMa)
to transport the collection
plate from a predefined
transfer position to any
destination on the robotic
worktable (plate reader,
hotels, autosampler, etc.)
for further analysis (UV
measurement, ELISA,
HPLC, MS, etc.)
• Assign labels (resin
name and column
types) by using
the plate layout
configurator
• Use the
neutralization option
to adjust the pH of
the fractions after
the acidic elution of
mAbs
Note: Confirm with
Tecan suitability
for any specific
application.
Calculate the precise
volume of your collected
fractions for subsequent
data plotting
Technical Support: 1-800-424-6723 • www.bio-rad.com 5
Table 3. Recommended equipment for automated handling of
Foresight RoboColumn units, continued.
Equipment Details Tips and tricks
Reagent and
sample
reservoirs
Collection plate Use any type of
A broad spectrum
of troughs, tubes,
and plates can be
positioned onto the
robotic worktable
by using the
appropriate Tecan
carrier
multi-well plate
which matches the
ANSI/SDS
standard (UV or
non-UV readable)
Use an 8-, 48- or 96deep well reservoir for
buffer
• Use a half area
96-well microplate to
collect fractions in a
volume range from
25–175 µl
• Use a standard 96well microplate to
collect fractions in a
volume range from
75–340 µl
• Use 96-deep well
reservoirs to collect
high volume fractions
up to 2.0 ml
3.2 Sample Preparation
Loading a clarified sample onto Foresight™ RoboColumn
units is
recommended because unclarified sample may cause clogging of
the top and bottom filter frits. This may impact the performance of
the column.
3.3 Experiment Setup
Foresight RoboColumn units are suitable for applications such as
parallel screening and optimization of chromatographic conditions
in downstream process development. The flexible design enables
arrangement of up to 12 rows (8 columns per row) of Foresight
RoboColumn units on a 96-well array plate (available upon request)
according to application’s requirements.
6 Technical Support: 1-800-424-6723 • www.bio-rad.com
When using the HT process development approach for Foresight
RoboColumn units, DoE is recommended for experiment setup,
as it enables efficient and parallel screening of many different
chromatographic parameters such as resin type, pH, conductivity
or ionic strength, and salt type.
For experiment setup and configuration of the Foresight
RoboColumn units, the Te-Chrom Wizard is recommended. See
Section 3.4.
3.4 Te-Chrom Wizard
The Te-Chrom Wizard is a dialog-based graphical user interface
offered by Tecan and based on the Freedom EVOware software.
The Te-Chrom Wizard was developed for use with Foresight
RoboColumn units and enables configuration of hardware, plate
layout, and setup of the chromatographic process, including
relevant parameters such as flow rate and volume, without direct
script writing (see Figure 1).
Please confirm with Tecan suitability for any specific application.
For more detailed information please refer to the Te-Chrom
software manual.
Fig. 1. Te-Chrom Wizard features.
Technical Support: 1-800-424-6723 • www.bio-rad.com 7
Section 4
2 x
Protocol
The protocol is designed as a general guideline for working with
Foresight™ RoboColumn units. Adjustment may be required
depending on type of application, sample, resin type, and
Foresight RoboColumn units dimensions.
One product package contains 8–96 individual prepacked
Foresight RoboColumn units arranged on a 96-well array plate,
and two removable cover seals for storage (see Figure 2).
Fig. 2. 96 Foresight RoboColumn units.
4.1 Detailed Protocol
1. Remove cover seals
a. First, gently peel off the
top seal from the columns
b. Next, gently peel off
the bottom seal from the
columns
8 Technical Support: 1-800-424-6723 • www.bio-rad.com
2. Remove excess storage
A
solution from needle adaptor
inlet
Remove remaining storage
solution in the top part of the
columns by blotting them
upside down on a soft paper
tissue
3. To adjust the Te-Chrom
module operating height,
press and hold the fastener
(C) and move the module to
the required height of the
scale (D)
Note: Make sure that the
columns are positioned as
close as possible to the
collection plate before starting
the experiment
4. Loading the Foresight
RoboColumn units in the
Te-Chrom module
a. Move the locking slider (A)
to the left mechanical stop
b. Load the columns (B)
c. Move the locking slider
(A) to the right mechanical
stop to lock the columns
Note: Make sure that all
columns are securely fixed
into the array plate before
starting the experiment.
Technical Support: 1-800-424-6723 • www.bio-rad.com 9
5. Experiment setup with the
Te-Chrom Wizard
a. Hardware configuration
• Select the appropriate
configuration of the
Te-Chrom module
• Check whether the
module is equipped with
a waste tray
• Define a transfer position
for the collection plate
b. Plate layout
• Select the RoboColumn
labware
• Define the required
RoboColumn matrix
• Select the RoboColumn
dimension
• Set a desired label such
as resin name
• Import or export the plate
layout
c. Process configuration
• Define the
chromatographic
workflow by setting
a desired number of
process steps
• Use the Continuous
Fraction Collection
option to collect fractions
over multiple plates, to
prevent gaps between
plates pipetted in different
process steps and to
optimize the consumption
of collection plates
10 Technical Support: 1-800-424-6723 • www.bio-rad.com
d. Configure your process
steps
• Defiine a process
step name such as
Equilibration
• Check the Process
Full Plate option if
appropriate:
i. When checked,
the full array will be
processed
ii. When not checked,
the array is processed
column by column
• Set parameters for
tip washing
• Set individual
chromatographic
parameters
i. Select the liquid
ii. Set the dispensing
volume
iii. Set the required liquid
class for aspiration
iv. Set the required
flow rate (cm/h)
Note: Calculate the
required residence
time (t) by dividing the
column volume (CV) by
the volumetric flow rate
(Uv). t = CV/U
v
Technical Support: 1-800-424-6723 • www.bio-rad.com 11
• Check the waste option
if appropriate:
i. When checked, the
waste tray will be
engaged by the
RoMa or Liquid
Handling Arm (LiHa)
ii. When not checked,
the flowthrough will
be collected in a
desired collection
plate
• Check the Neutralization
option to perform a pH
adjustment, for example
after acidic elution of
mAbs
• Check the Data
Collection option to
evaluate the collection
plate in a required reader
device such as a UV
reader
• Select the number of
fractions according to the
given dispensing volume
for the chromatography
step if the waste option is
not checked
e. Plate handling
• Add the required amount
of source plates
• Add the required amount
of destination plates
12 Technical Support: 1-800-424-6723 • www.bio-rad.com
After the experiment setup the Te-Chrom Wizard will generate a
chromatographic method in Freedom EVOware software automatically.
f. Reporting
• Set output folder
• Select output format
6. Run chromatographic method
Note: Make sure all required
solvents and consumables
such as collection plates are in
their designated positions on
the robotic worktable before
running the chromatographic
method.
7. Data evaluation
a. Determine the precise
volume of the collected
fractions by using an
appropriate plate reader
b. Determine the protein
concentration by
measuring UV
Note: After path length
detection with the RoMa,
transfer the collection
plate to any location on
the robotic worktable, or
to a secondary device
for further analysis such
as HPLC, MS, ELISA, or
CGE/SDS-Page.
Technical Support: 1-800-424-6723 • www.bio-rad.com 13
8. Data plotting
Plot the UV data against the
fraction volume to create a
pseudo-chromatogram
Note: Make sure that blank
values of the used solvents
are subtracted from the UV
data before plotting.
9. Regeneration and storage
a. For appropriate
regeneration of Foresight
RoboColumn units please
refer to
www.bio-rad.com/process
for the recommendation of
the specific resin
b. For appropriate storage of
Foresight RoboColumn
units, refer to Table 1
14 Technical Support: 1-800-424-6723 • www.bio-rad.com
Section 5
Troubleshooting Guide
Fault Possible cause Action
Foresight™
RoboColumn
units are
clogged
Foresight
RoboColumn
units are leaking
• The sample is too
viscous
• There is too much
cell debris in the
sample
• The column has
been regenerated
and used for too
many cycles
• Columns are
clogged
• The O-ring seal is
damaged
• The insertion depth
of the robotic needle
is incorrect
• Increase dilution of
• Centrifuge and/or
• Reduce the number
• See above
• Check the taught
• Check the insertion
the cell isolate before
lysis, or dilute after
the lysation
filter the sample if
unclarified sample
has been used
of cycles
X and Y coordinates
for RoboColumn
labware for precision
depth (16 mm
from the top of
the column) of the
robotic needle
Foresight
RoboColumn
units are lifted
up from the
96-well array
plate during the
pull-out of the
robotic needles
Low protein
recovery in the
elution fraction(s)
Columns are not securely
seated into the 96-well
array plate
The residence time for
sample loading is too low
Technical Support: 1-800-424-6723 • www.bio-rad.com 15
Make sure that all
columns are securely
seated into the
96-well array plate by
firmly pressing down
before starting the
experiment
Decrease the flow rate or
increase residence time
for sample loading
Section 6
Ordering Information
Foresight RoboColumn units 200 and 600 µl
Product Mode Catalog #
Foresight™ Nuvia™ S
RoboColumn units, 200 µl
Foresight Nuvia S
RoboColumn units, 600 µl
Foresight Nuvia Q
RoboColumn units, 200 µl
Foresight Nuvia Q
RoboColumn units, 600 µl
Foresight™ Nuvia™ cPrime
RoboColumn units, 200 µl
Foresight Nuvia cPrime
RoboColumn units, 600 µl
Foresight™ UNOsphere™ S
RoboColumn units, 200 µl
Foresight UNOsphere S
RoboColumn units, 600 µl
Foresight UNOsphere rS
RoboColumn units, 200 µl
Foresight UNOsphere rS
RoboColumn units, 600 µl
Foresight UNOsphere Q
RoboColumn units, 200 µl
Foresight UNOsphere Q
RoboColumn units, 600 µl
Foresight™ CHT™ Type I,
40 µm RoboColumn units,
200 µl
Foresight CHT Type I,
40 µm RoboColumn units,
600 µl
Foresight CHT Type II,
40 µm RoboColumn units,
200 µl
Foresight CHT Type II,
40 µm RoboColumn units,
600 µl
Strong Cation 732-4801
Strong Cation 732-4802
Strong Anion 732-4804
Strong Anion 732-4805
™
Mixed Mode
(Hydrophobic-Cationic)
Mixed Mode
(Hydrophobic-Cationic)
Strong Cation 732-4813
Strong Cation 732-4814
Strong Cation 732-4816
Strong Cation 732-4817
Strong Anion 732-4819
Strong Anion 732-4820
Mixed Mode
(Metal Affinity-Cationic)
Mixed Mode
(Metal Affinity-Cationic)
Mixed Mode
(Metal Affinity-Cationic)
Mixed Mode
(Metal Affinity-Cationic)
732-4807
732-4808
732-4822
732-4823
732-4825
732-4826
16 Technical Support: 1-800-424-6723 • www.bio-rad.com
Foresight™ Chromatography Media Filter Plates
Product Mode Catalog #
Foresight™ Nuvia™ S
Filter Plate, 20 µl
Foresight Nuvia Q
Filter Plate, 20 µl
Foresight™ Nuvia™ cPrime™
Filter Plate, 20 µl
Foresight™ UNOsphere™ S
Filter Plate, 20 µl
Foresight UNOsphere rS
Filter Plate, 20 µl
Foresight UNOsphere Q
Filter Plate, 20 µl
Foresight™ CHT™ Type I,
40 µm Filter Plate, 20 µl
Foresight CHT Type II,
40 µm Filter Plate, 20 µl
Strong Cation 732-4701
Strong Anion 732-4703
Mixed Mode
(Hydrophobic-Cationic)
Strong Cation 732-4710
Strong Cation 732-4712
Strong Anion 732-4714
Mixed Mode
(Metal Affinity-Cationic)
Mixed Mode
(Metal Affinity-Cationic)
732-4705
732-4716
732-4718
Technical Support: 1-800-424-6723 • www.bio-rad.com 17
Foresight™ Chromatography Media Columns
Product Mode Catalog #
Foresight™ Nuvia™ S
Column, 1 ml
Foresight Nuvia S
Column, 5 ml
Foresight Nuvia Q
Column, 1 ml
Foresight Nuvia Q
Column, 5 ml
Foresight™ Nuvia™ cPrime™
Column, 1 ml
Foresight Nuvia cPrime
Column, 5 ml
Foresight™ UNOsphere™ S
Column, 1 ml
Foresight UNOsphere S
Column, 5 ml
Foresight UNOsphere rS
Column, 1 ml
Foresight UNOsphere rS
Column, 5 ml
Foresight UNOsphere Q
Column, 1 ml
Foresight UNOsphere Q
Column, 5 ml
Foresight™ CHT™ Type I,
40 µm Column, 1 ml
Foresight CHT Type I,
40 µm Column, 5 ml
Foresight CHT Type II,
40 µm Column, 1 ml
Foresight CHT Type II,
40 µm Column, 5 ml
Strong Cation 732-4720
Strong Cation 732-4740
Strong Anion 732-4721
Strong Anion 732-4741
Mixed Mode
(Hydrophobic-Cationic)
Mixed Mode
(Hydrophobic-Cationic)
732-4722
732-4742
Strong Cation 732-4730
Strong Cation 732-4750
Strong Cation 732-4731
Strong Cation 732-4751
Strong Anion 732-4732
Strong Anion 732-4752
Mixed Mode
(Metal Affinity-Cationic)
Mixed Mode
(Metal Affinity-Cationic)
Mixed Mode
(Metal Affinity-Cationic)
Mixed Mode
(Metal Affinity-Cationic)
732-4735
732-4755
732-4736
732-4756
18 Technical Support: 1-800-424-6723 • www.bio-rad.com
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