PROCESSSkillPak™ 5 mL Instruction Manual for TOYOPEARL®,
TSKgel® and Ca++Pure-HA™ Process MediaIntroduction
SkillPak 5 mL columns (8 mm ID × 10 cm) are designed for fast method
development or resin screening and are pre-packed with TOYOPEARL, TSKgelor Ca++Pure-HA process chromatography media for bioseparations. Thesecolumns are designed to be operated with commonly used low or medium
pressure liquid chromatography systems.
SkillPak 5 mL columns are reproducibly packed and take into account thevarying compressibility of each resin. This provides an accurate representationof conditions found in full scale columns.
Product Description
Hardware Materials

The hardware of SkillPak 5 mL columns is composed of bio-compatible
polymers, mainly polypropylene. Table 1 lists properties and operation
specifications of the SkillPak 5 mL columns.
Stop plug
End cap

Column tube
End cap
Stop plugInstructions for Use
Installation

SkillPak 5 mL columns are ready to use upon receipt. Connect the column tothe instrument following the recommended flow direction as indicated on thecolumn label. Connect inlet/outlet of column to instrument using 10-32 fitting.
Finger-tight is recommended.
Equilibration

Before the first sample injection, the column should be equilibrated with at least
10 CV of the respective loading buffer. See Table 1 for standard flow rate andmaximum operating pressure. Slowly increase flow until it reaches the desired
running flow rate. Do not exceed the maximum flow rate.
Buffer composition and gradient design is dependent on the resin type and resin
chemistry. Please refer to the TOYOPEARL, TSKgel or Ca++Pure-HA productdocumentation for more information about recommended mobile phases.
Table 1. Specifications of SkillPak 5 mL columns
Column dimension

8 mm ID × 10 cm bed height

Volume

5 mLStandard flow rate

1.3 mL/min (150 cm/hr)

Maximum flow rate

5 mL/min (600 cm/hr) for TOYOPEARL M andC grade resins; 2.5 mL/min (300 cm/hr) for
TOYOPEARL S-grade, TSKgel, Ca++Pure-HA andTOYOPEARL F grade resins, including TOYOPEARL
AF-rProtein A HC-650F, TOYOPEARL AF-rProtein
L-650F, and Ca++Pure-HA

Maximum operating pressure

0.3 MPa for TOYOPEARL resins, ≤ 0.4 MPa for
TSKgel resins and Ca++Pure-HAConnectionsStandard fittings (10-32 for 1/16 inch capillary)

Shipping buffer

20% ethanol for TOYOPEARL and TSKgel, 20 mmol/L
phosphate with 20% ethanol for Ca++Pure-HA

Asymmetry factor (As)specifications

0.8-1.4 for TOYOPEARL and TSKgel, 0.8-2.6 for
Ca++Pure-HA

Column performance test

The chromatographic performance of a SkillPak 5 mL column should be tested
initially and at regular intervals and is determined by the asymmetry factor.
The specification can be found in Table 1.
For most of the TOYOPEARL S-grade, F-grade, M-grade, and TSKgel resins,the evaluation can be carried out as described below:• Equilibrate columns with ≥ 10 CV of 150 mmol/L NaCl @ ≤ 300 cm/hr• Decrease the flow rate to 150 cm/hr and inject 1% CV of 3 mol/L NaCl tracer• Wash with 2 CV of 150 mmol/L NaCl @ 150 cm/hr		Note: the response can be detected using the conductivity meter.
For TOYOPEARL GigaCap series of resins, the evaluation can be carried out
as described below:• Equilibrate columns with ≥ 10 CV of 1 mol/L NaCl @ ≤ 300 cm/hr• Decrease the flow rate to 150 cm/hr and inject 1% CV of 3 mol/L NaCl tracer• Wash with 2 CV of 150 mmol/L NaCl @ 150 cm/hr		Note: the response can be detected using the conductivity meter.
For Ca++Pure-HA media, the evaluation can be carried out as described below:• Equilibrate columns with ≥ 10 CV of 0.2 mol/L sodium phosphate,		 pH 6.8 @ ≤ 300 cm/hr• Decrease the flow rate to 150 cm/hr• Inject 1% CV of 1% acetone tracer in 0.2 mol/L sodium phosphate,		 pH 6.8 @ 150 cm/hr		Note: the response can be detected using UV @ 280 nm.
Regeneration and storage conditions

SkillPak 5 mL columns can be washed/regenerated. The columns can also
be returned to shipping buffer for later use. Please refer to the TOYOPEARL,TSKgel or Ca++Pure-HA product documentation for more information aboutstorage and cleaning solutions.
SkillPak 5 mL columns are reusable as long as the resin remains in acceptablecondition, which will largely depend on the care taken, the cleaning performed
and the tolerance of the resin to applied conditions. The columns are notdesigned to be repacked.

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Tosoh Bioscience, TSKgel and TOYOPEARL are registered trademarks of Tosoh Corporation.
Ca++Pure-HA is a registered trademark of Tosoh Bioscience LLC in the USA, EU, India, and Japan.
SkillPak is a registered trademark of Tosoh Bioscience LLC in the USA.

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