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Tween® 80 & Tween 20

NOTE S

Analysis of MAb 221 in Tween 80 or Tween 20 with Sepax SEC Columns

Column: SRTTM SEC-300 (5um, 4.6x300mm) Mobile phase: 150 mM phosphate buffer, pH 7.0 Flow rate: 0.35 mL/min; Wavelength: 214 nm Temperature: RT; Injection volume: 3 µL Column pressure: 560 psi Sample: MAb 221 (1.0 mg/mL) MAb 221 (1.0 mg/mL) in 0.1%Tween 80

THE CHALLENGE:

To prevent a Tween 80 detergent peak just where one's protein elutes. Commonly used surfactants, such as Tween 80 (Polysorbate 80) or Tween 20 (Polysorbate 20) in drug formulation buffers, can co-elute with the main protein peak on SEC columns.

THE SOLUTION:

Sepax SEC: (SRT 5um or Zenix 3um)

Delivers unrivaled, high resolution separations, with virtually no Tween 80 or Tween 20 interference, due to a unique, proprietary, surface coating technology. Sepax SEC allows one to focus on peaks of interest. SRT SEC-300, 5um silica based SEC, offers high resolution and non-interfering separations of MAb 221 with or without the presence of 0.1% Tween 80 or Tween 20.

Distributed By

The Nest Group, Inc.

45 Valley Road Southborough, MA 01772 USA Tel: (508) 481-6223 | Fax: (508) 485-5736

Column: SRT SEC-300 (5um, 300A, 4.6x300mm) Mobile phase: 150 mM phosphate buffer, pH 7.0; Flow rate: 0.35 mL/min; Detector: Refractive Index; Temperature: RT; Sample: mAb 221 (1.4 mg/mL); Injection Volume: 3 µL

E-mail: [email protected] www.nestgrp.com

Toll Free: 1-800-347-6378

Tween 80 & Tween 20

NOTE S

Tween 80 and Tween 20 have no impact on SRT SEC mAb separations

Data Supported Facts: :

As shown at right, mixtures of MAb with Tween 80 (0.1%) and MAb with Tween 20 (0.1%) were subject to size-exclusion chromatography on SRT SEC-300 (5um, 300A, 4.6x300mm) and detected using Refractive Index. Furthermore, water, Tween 80 and Tween 20 samples were separately injected without the use of the SRTTM SEC column. The consistency of these three separation profiles suggests Tween 80 or Tween 20 samples pass through the column with no impact on the separation of a mAb.

*No column: Directly injected H2O, Tween 80 or Tween 20 into the HPLC system without a column connected.

Column Condition:

Column: SRT SEC-300 (5um, 300A, 4.6x300mm); Mobile phase: 150 mM Phosphate Buffer, pH=7.0; Flow rate: 0.35 mL/min; Detector: Refractive Index; Temperature: RT; Sample: MAb 221 (1.4 mg/mL); Injection Volume: 3 µ L

Tween 20 & Tween 80 are registered trademarks of ICI Americas, Inc.

Ordering Information:

Distributed By

The Nest Group, Inc.

45 Valley Road Southborough, MA 01772 USA Tel: (508) 481-6223 | Fax: (508) 485-5736

T 215300-7830 T 215300-4630 T 213300-7830

SRT SEC-300, 5um, 300A, 7.8x300mm SRT SEC-300, 5um, 300A, 4.6x300mm Zenix SEC-300, 3um, 300A, 7.8x300mm

E-mail: [email protected] www.nestgrp.com Toll Free: 1-800-347-6378

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Sepax Applications-Tween.doc