Bio-Rad Next Generation

Medium Pressure Liquid Chromatography

Instruments
 Chromatography - INTRODUCTION
Chromatography is the science of separating bio-molecules by taking advantage of how they interact with stationary phase.

 Column based chromatography method is the most effective way to isolate proteins

 A simple set up such as a gravity column or syringe column can be used

Chromatography instruments provide reproducibility through automations

 A pump adds speed, better gradients and reproducibility

 A sample injection valve adds automation and consistent application volumes

 A detector monitor the separation and provides the chromatogram record of the run

 The fraction collector ensures that discrete volumes will be collected automatically

Bio-Rad Provides a range of chromatography columns and equipments

 Empty columns such as Econo-Column (and more)

 Prepacked columns such as BioScale Mini and High-Resolution Enrich series (and more)

 Simple systems like the Econo System

 More versatile systems like Biologic LP, Duoflow and NGC Series
 INTRODUCTION – NGC Series Systems

 NGC Medium-Pressure Chromatography Systems fit your throughput requirements when purifying biomolecules for analytical- and
preparative-scale applications.

 Each system is composed of interchangeable modules that confer different system capabilities and available in 10ml and 100ml system
pumps.

NGC Quest™ and NGC™ Quest Plus Systems

 Separating biomolecules at high resolution using accurate gradients

NGC Scout™ and NGC™ Scout Plus Systems

 Medium throughput applications requiring rapid scouting of protein purification conditions

with automated gradients and buffer preparation

NGC Discover™ and NGC™ Discover Pro Systems

 Higher throughput applications with rapid, robust automation and a need for method development
 NGC 100 FPLC Systems and Configurations

Product NGC Quest NGC Quest NGC NGC Scout100 NGC Discover NGC Discover
Description 100 100 Plus Scout100 Plus 100 100 Pro
F100 Pump Module ● ● ● ● ● ●

Mixer Module ● ● ● ● ● ●

Sample inject valve ● ● ● ● ● ●

module
Single wavelength ● - ● - - -
detector module
Multi wavelength - ● - ● ● ●
detector module
Buffer Blending valve - - ● ● ● ●
module
pH valve module - - ● ● ● ●

Sample pump module - - - - ● ●

Inlet valve module - - - - ● ●

Column switching - - - - ● ●
valve module 100ml
Outlet Valve Module - - - - - ●
 NGC 10 FPLC Systems and Configurations

Product NGC Quest 10 NGC Quest 10 NGC Scout10 NGC Scout10 NGC Discover NGC Discover
Description Plus Plus 10 10 Pro
F10 Pump Module ● ● ● ● ● ●

Mixer Module ● ● ● ● ● ●

Sample inject valve ● ● ● ● ● ●

module
Single wavelength ● - ● - - -
detector module
Multi wavelength - ● - ● ● ●
detector module
Buffer Blending valve - - ● ● ● ●
module
pH valve module - - ● ● ● ●

Sample pump module - - - - ● ●

Inlet valve module - - - - ● ●

Column switching - - - - ● ●
valve module 100ml
Outlet Valve Module - - - - - ●
 NGC FPLC Systems and Modules Specifications

System Pumps 100 ml/min system pump kit with the necessary tubing
and fittings, for creating buffer gradients

Pump type Reciprocating piston

Flow rate 10 ml/min pumps: 0.001–10 ml/min (normal range)

setting 100 ml/min pumps: 0.01–100 ml/min (normal range

±2% (conditions: 0.1–10 ml/min, pressure

Flow rate
<600 psi (4.1 MPa, 41 bar),
accuracy
viscosity: 0.5–3.7 cP

Pressure 10 ml/min pumps: 0–25.2 MPa (3,650 psi)

range 100 ml/min pumps: 0–10 MPa (1,450 psi) Use the F100 NGC System Pumps for producing automated proportioned
solutions and flow rates of up to 100 ml/min at 1,450 psi (100 bar, 10
Viscosity
0.5–10.8 cP (for 10 ml/min and 100 ml/min pumps) MPa).
range
Features and Benefits of the NGC F100 Chromatography System Pump
Module
 Enables different flow rates (up to 100 ml/min) when used with the
buffer Blending valve Can be controlled to avoid overpressure
 NGC FPLC Systems and Modules Specifications - Sample Pump Module

100 ml/min sample pump kit with the necessary tubing

and fittings, for automated large-volume sample
application The NGC Sample Pump Module, when used with the Sample Inject valve
enables the injection of large sample volumes and allows repetitive, accurate,
and multiple loadings from a single sample stock.
Features and Benefits of NGC Chromatography Sample Pumps
 Eliminate the risk of contaminating system pumps
 Allow automated loading of large sample volumes, either directly to the
column or via a DynaLoop™ Sample Loop, at a flow rate of up to 100 ml/min
at 1,450 psi
 Feature an integrated pressure sensor that protects column and media from

Pump type Piston pump, metering type overpressure

Flow rate  Automate sample addition, thus increasing accuracy between experimental
0.01–100 ml/min
setting runs
Flow rate
±2%
accuracy
Pressure range 0–10 MPa (1,450 psi)
Viscosity range 0.5–10 cP
 NGC FPLC Systems and Modules Specifications - Mixer Module

 The NGC Mixer Module homogenizes buffers from the two system pumps (A and B) and includes a
mixer motor assembly and an integrated system pressure sensor. Overall, efficient gradient mixing
10 ml/min kit with the necessary tubing and
at higher flow rates can be achieved.
fittings, upgradable for use with F100 systems
 This mixer module comes with the 263µl barrel, for use with NGC 10 Systems, and can be
extended with small mixer barrels of increasing size ranging from 750µl and 2ml.
 This mixer module is also compatible with NGC 100 Systems through the use of an F100 mixer (750
µl), which can be extended with 2ml, 5ml or 12ml large mixer barrels.
Mixing principle: Chamber with magnetic stirrer
Mixer volume: 263 µl (included), 750 µl (included), 2 ml, 5 ml (for F10 pumps)
750 µl (included), 2 ml (included), 5 ml, 12 ml (for F100 pumps)
Gradient composition accuracy: ±0.5% (conditions: 3–97% B, 0.25–10 ml/min for F10 pumps)
±0.5% (conditions: 3–97% B, 0.25–100 ml/min for F100 pumps)
Features and Benefits of NGC Chromatography Mixer Modules
 Integrated pressure sensor
 Interchangeable mixing chambers for efficient gradient mixing at higher flow rates
 NGC FPLC Systems and Modules Specifications - Sample Inject valve Module

Automated application of small sample

volume samples

 The NGC Sample Inject Valve enables the system to load a specific predetermined volume of sample

onto a column.

 This is useful when applying small sample volumes manually via sample loops or large volume samples

using a sample pump. The sample inject valve has a maximum operating pressure of 3,650 psi.

Features and Benefits of the NGC Chromatography Sample Inject Valve Module

 Enables the loading of a specific predetermined volume of sample onto a column

 NGC FPLC Systems and Modules Specifications - Inlet Valve Module

For automated switching between multiple buffers and Inlet A 8 inlets

samples during method development Inlet B 8 inlets
Sample inlet* Up to 16 inlets

 The NGC Inlet Valve Module allows inline buffer preparation and pH

gradient generation for quick pH scouting.

 It speeds up method development by allowing the automatic

selection of buffers and cleaning solution across multiple runs on

NGC™ Chromatography Systems.

 NGC FPLC Systems and Modules Specifications
UV & UV/Vis Detectors

Single-wavelength: 255 nm (nucleic acids) or 280 nm

Wavelength (proteins)
Multi-wavelength (up to 4): 190–800 nm

Absorbance range 0 to >2.8 Au

Linearity 0–2 Au within ±5%

1,450 psi (10 MPa) for 5, 10 mm flow cells

Operating pressure
700 psi (5 MPa) for 2 mm flow cells

Flow cells
Operational
Preparative: 2 mm (cell volume: 20 µl)
interchangeable flow
Analytical: 5 mm (cell volume: 9 µl)
cells available in path
Analytical: 10 mm (cell volume: 18 µl)
lengths of 2, 5, and
10 mm
 NGC Discover Pro: Multi wavelenth Module

For protein, peptide, nucleic acid, and

chromophore detection  The NGC Multi-Wavelength Detector adds flexibility to your chromatography system, enabling the
simultaneous monitoring of four wavelengths between 190 and 800 nm for the detection of
proteins, peptides, nucleic acids, and chromophores such as hemoglobin in complex biological
mixtures.
 The NGC UV/Vis Detector Module features an integrated conductivity monitor to measure buffer
conductivity between 0.01–999 mS/cm using a nominal volume of 6 µl. This detector can be used
with three interchangeable flow cells with path lengths of 2, 5, and 10 mm.
 A flow restrictor can be added to the flow path to generate a backpressure that prevents the
formation of air bubbles in the UV flow cells.

Features and Benefits of the NGC Chromatography Multi-Wavelength Detector Module

 Multi-wavelength UV/Vis detector for simultaneously monitoring wavelengths between 190–800
nm for the detection of proteins, peptides, nucleic acids, and chromophores
 Can be used with two interchangeable flow cells with path lengths of 2, 5, and 10 mm
 The two flow cells can be used on both the single- and multi-wavelength detectors
 NGC FPLC Systems and Modules Specifications
pH Monitor
Conductivity Monitor
pH reading
0–14
Conductivity reading range 0.01 µS/cm–999 mS/cm range

Accuracy ±2% ±0.1 pH unit within pH

Accuracy
2–12
Operating pressure 0–5.5 MPa (800 psi)
0–70 psi with pH probe
Flow cell volume 6 µl
Operating inline
Temperature monitor range 4–100°C pressure and 0–500 psi in bypass
Temperature monitor accuracy ±2% mode

Flow cell 100 µl (210 µl including

volume internal flow paths)
 NGC FPLC Systems and Modules Specifications - Column Switching Valve Module

 Use the NGC Column Switching Valve 100 ml to connect up to 5 columns to your NGC 100
Accommodate up to 5 columns, for column scouting
Chromatography System.
and reverse flow applications using NGC 100
Chromatography Systems
 This valve has an internal bypass mode that enables buffers to bypass the connected columns
when priming or cleaning the system. The valve also has the ability to reverse the flow, which is
ideal for column cleaning and certain applications.
 The column switching valve has one inlet port, one outlet port, and five pairs of column
connection ports. The column position can be changed manually or automatically within
Chrome lab software.
Features and Benefits of the NGC Column Switching Valve Module 100 ml
 Has 1 inlet port, 1 outlet port, and 5 pairs of column connection ports
 Enables quick and easy column scouting without the need to re-plumb
 Includes integrated pressure sensors, which measure pre- and delta-column pressures and
protect columns and media from overpressure
 Features internal bypass mode, allowing buffers to bypass the connected columns when
priming or cleaning the system
 Provides reverse flow capability, ideal for column cleaning
 Up to 3 column switching valves can be used, allowing up to 15 columns to be connected
 NGC FPLC Systems and Modules Specifications - Outlet Valve Module

 Use the NGC Outlet Valve either as a stand-alone unit or in conjunction with the BioFrac™
Automated fraction collection of large-
volume fractions with up to 12 vessels Fraction Collector

 As a single, stand-alone unit, the outlet valve is capable of collecting up to 12 large-volume

fractions such as unbound flow material or waste. Two outlet valves may be daisy-chained to

collect up to 23 large-volume fractions, including waste.

 When a BioFrac Fraction Collector is connected the collection sequence can be programmed to

collect smaller-volume fractions in the BioFrac Fraction Collector and larger-volume fractions

via the outlet valves, providing full flexibility.

Features and Benefits of the NGC Outlet Valve Module

 Collect up to 12 large-volume fractions with a single valve

 Collect up to 23 large-volume fractions with two valves

 Collect variable fraction sizes with the BioFrac Fraction Collector

 NGC FPLC Systems and Modules Specifications - Buffer Blending valve Module

For inline buffer preparation and pH

 The NGC Buffer Blending Valve is used for automatic inline buffer preparation and generation of pH
gradient generation for quick pH scouting
gradients for quick pH scouting.
 When used in conjunction with the dual gradient pump system, the buffer blending valve enables
automated elution gradients and highly accurate buffer blending for rapid scouting and method
development.
 In buffer blending mode, acid, base, water, and salt are blended together in the required proportions to
form a buffer of specified pH over a linear salt gradient or a pH gradient at a specified salt
concentration.
 In addition to buffer blending, this valve can also be used to double the flow rate of a salt gradient to
20 ml/min on an F10 pump.
 A flow rate of up to 175 ml/min can be achieved with the F100 pump.
Features and Benefits of the NGC Chromatography Buffer Blending Valve Module
 Can be used to double the flow rate of a salt gradient to 20 ml/min on an F10 pump and to 175 ml/min
on an F100 pump
 For automatic inline buffer preparation and generation of pH gradients for quick pH scouting
 NGC FPLC Systems and Modules Specifications

Air Sensors

Up to 8 total air sensors (1 for end of sample detection,

Number of sensors
remaining are for end of buffer detection)

Accuracy ±0.1 pH unit within pH 2–12

Placement of built-in sensors End of buffer, end of sample

Sensing principle Acoustic

NGC Quest and Quest Plus Instruments

NGC Quest™ Chromatography System

Includes 10 ml or 100ml/min pumps, single-wavelength (UV)

and conductivity detection, and automated sample
injection, for the purification of proteins.

NGC Quest™ Plus Chromatography System

Includes 10 ml or 100ml/min pumps, multi-wavelength

(UV/Vis) and conductivity detection, and automated
sample injection, for the purification of proteins
NGC Scout and Scout Plus

Features and Benefits of the NGC Scout Chromatography System

 A flexible, customizable chromatography system to suit both your application
and workflow needs:
 The Guided Fluidics selection tool (patent pending) coupled with the Module
Discovery system enables a single-click application-based system setup
through automated module function recognition
 The Tier Rotate™ design allows optimal placement of valves and detectors to
minimize the flow path
 Compact footprint – fits in a refrigerator
 Point-to-Plumb™ lighting shows the step-by-step LED-guided plumbing of the
system
 Unique ChromLab software provides powerful system control, automated
methods, and analysis options
 NGC Discover and Discover Pro

For simultaneously detecting proteins, peptides, nucleic acids, and chromogenic

molecules using expanded scouting options.
Features and Benefits of the NGC Discover 100 Pro System
 Automated 100 ml/min pumps provide accurate gradients for high-resolution
separation of biomolecules
 Automated injection of large sample volumes using fixed or dynamic loops
 Gradient separations at different pH values and buffer pH monitoring, for
rapid method development
 Automated buffer and salt gradients with buffer blending during automated
inline buffer preparation at flow rates of up to 200 ml/min, for rapid scouting
of purification conditions

Includes 100 ml/min pumps, multi-  Simultaneous four-wavelength detection of proteins, peptides, nucleic acids,

wavelength (UV/Vis) and conductivity and chromophores, with conductivity measurement and pH detection

detection, expanded scouting options, large-  Fraction collection in combination with large-volume collection using a 12-

sample injection, outlet valve and column port outlet valve

switching of up to 5 columns, for method  Compatibility with the BioFrac™ Fraction Collector for sample collection

development
BioFrac™ Fraction Collector

 The BioFrac Fraction Collector is ideal for analytical and preparative

chromatography applications; it's easy to program and can be used for basic
or complex fraction collection schemes at flow rates ≤100 ml/min.

 Available as a stand-alone fraction collector or as a component of the NGC,

BioLogic DuoFlow™ and BioLogic™ LP Chromatography Systems, the BioFrac
Fraction Collector offers flexibility with numerous collection rack options,
including both custom and off-the-shelf collection racks.

 The custom racks provide a wide range of collection capability, from drops to
liters. Off-the-shelf racks extend the versatility of collection schemes and
provide cost-effective storage of samples. Off-the-shelf racks are
autoclavable, can be easily assembled, and lie flat, using little storage space.
 ChromLab™ Software

Features and Benefits of ChromLab Software

It controls all functions for laboratory-scale protein
 Use of the touch screen allows the same level of flexibility without the need for a
purification including instrument setup and
computer adjacent to the system, minimizing the total footprint
calibration, method development, real-time  Customizable chromatogram layouts with a variety of viewing and data analysis options
monitoring and system control, chromatogram  Rapid creation of trace comparisons allows easy analysis across multiple purification runs
comparison, and peak analysis.  Single-click peak detection and integration for quicker results
 Advanced integration parameters enable manual integration functions and peak addition
or removal for fine-tuning your data
 Intuitive, grouped data. Tabular layouts organize data for easy navigation
Applications and Uses of ChromLab Software
 Controls all functions of the NGC Medium-Pressure Chromatography System for lab-scale
protein purification
 Instrument control
 Method development to identify best methods
 Real-time monitoring of proteins, peptides, nucleic acids, and chromophores
 Chromatogram comparison and peak analysis