Spectra Manager BeStSel

High estimation accuracy fora wide range of proteins
including β-structure-rich-proteins such as antibodies.

We also offer CFR version
that complies with thedata management standardsof 21 CFR Part 11.

Providing eight types ofsecondary structure
information

Secondary structure

Helix 1 – regular α-helix
The middle part of α-helices

Helix 2 – distorted α-helix

Left-twisted

2-2 residues at the ends of α-helices

Anti 1 – left-twisted β-strand
Left-hand twisted antiparallel β-sheet

Anti 2 – relaxed β-strand

RelaxedRelaxed (slightly right-hand twisted) antiparallel β-sheet

Anti 3 – right-twisted β-strand
Right-hand twisted antiparallel β-sheet

Parallel β-strand
Parallel β-sheet

Right-twisted

TurnTurn, as defined by DSSPOthers3,10-helix, π-helix, β-bridge, bend, loop/irregularand invisible regions of the structure

Secondary structure information obtained by BeStSel algorism

Automatic Y-axis conversion and concentration calculationby amino acid sequence and absorption

APPLICATION Secondary structure prediction of therapeutic antibodies
CD spectra of antibodies(Eötvös Loránd University, Hungary).

thersThis program utilizes ‘BeStSel engine’, which is an algorism developed by Dr. József Kardos and Dr. András Micsonai, et. al. at ELTEOnTurPara

23ti
AntitiAn....

* HERCEPTIN is a trademark of Genentech, Inc.

102250An

Wavelength/nm

240

ix

230ix

220

10el

210

NRMSD
= 0.0220el

HERCEPTIN *0200

30NRMSD= 0.01

Experimental
FittedSD

190

RIABNI(Amgen Inc.)

05000

401Experimental
FittedSD

NRMSD= 0.02H

MRE/deg.cm2dmol-15000- MABTHERA- RIABNI- HERCEPTIN(F. Hoffmann-La Roche AG)

0H

MABTHRAExperimental
FittedSDFraction(%)

5000

Secondary structure prediction50

Evaluation of the Structural Stability of Antibody Drugs
Here, we report the results of a comprehensive analysis of the secondary structure of antibody
drugs under various pH conditions utilizing a seamless workflow that combines the automated
CD/Absorbance(Abs)/Fluorescence(FL) measurement system HTCD Plus and Spectra Manager
BeStSel.
Results of SSE showed that both Herceptin and RIABNI retained their secondary structuredown to pH 3, but the ratio of Helix, β-strand, and Others changed significantly at pH 2.
Spectra Manager BeStSel is a powerful tool for structural analysis of a wide range of proteins, HTCD Plus
High-Throughput Circular Dichroism Measurement System
including antibody drugs.

3000

pH2220

240

250

200

210

220

230

Wavelength / nm

CD spectrum of Trastuzumab

CD spectrum of Rituximab

Helix

β-strand4Turn

240

250

Others62
100–5–10
2 3 4 5 6 7 8 9 1020–2–4

2 3 4 5 6 7 8 9 10pHpH

Δ Fraction (%)Δ Fraction (%)

3–1

230

pH2

Wavelength / nm

Δ Fraction (%)

4210

pH10MRE/deg. cm2dmol–1

MRE/deg. cm2dmol–1

pH10200

Δ Fraction (%)

3000

420–2

2 3 4 5 6 7 8 9 10pH

2 3 4 5 6 7 8 9 10

pH

Secondary structure analysis results of Trastuzumab

Helix

β-strandOthers

Turn
80–5

2

Δ Fraction (%)Δ Fraction (%)

2Δ Fraction (%)

04

Δ Fraction (%)

40–2–10
2 3 4 5 6 7 8 9 10pH

2 3 4 5 6 7 8 9 10–4

pH

6420

2 3 4 5 6 7 8 9 10pH

2 3 4 5 6 7 8 9 10pH

Secondary structure analysis results of Rituximab

The contents of this material are for reference and illustrative
purposes only. Information, descriptions, and specifications in thispublication are subject to change without notice. JASCO assumes noresponsibility and will not be liable for any errors or omissions
contained herein or for incidental, consequential damages or losses inconnection with the furnishing, performance or use of this material.
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ISO-14001

CERTIFIED

Products described herein aredesigned and manufactured byISO-9001 and ISO-14001 certified
JASCO CorporationJASCO CORPORATION

2967-5, Ishikawa-machi, Hachioji-shi, Tokyo 192-8537 Japan
Tel: +81-42-649-5177 Fax:+81-42-646-4515
Web: www.jasco-global.com
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