Amgen Inc. v. F. Hoffmann-LaRoche LTD et al

Filing 728

MEMORANDUM in Support re #727 MOTION for Leave to File Under Seal Documents Containing Defendants' Trade Secrets Submitted in Connection with the Pendleton and Galvin Declarations filed by F. Hoffmann-LaRoche LTD, Roche Diagnostics GmbH, Hoffmann LaRoche Inc.. (Attachments: #1 Exhibit A#2 Exhibit B#3 Exhibit C)(Toms, Keith)

Amgen Inc. v. F. Hoffmann-LaRoche LTD et al

Doc. 728 Att. 1

Case 1:05-cv-12237-WGY

Document 728-2

Filed 07/16/2007

Page 1 of 2

UNITED STATES DISTRICT COURT DISTRICT OF MASSACHUSETTS

AMGEN INC., )

) ) ) )

v. )

Plaintiff, )

I Exhibit A

C. A. No.: 05-CV-I2237-WGY

F. HOFFMANN-LA ROCHE LTD., )

a Swiss Company, ROCHE )

DIAGNOSTICS GmbH, a German )
Company and HOFFMANN-LA ROCHE )
INC., a New Jersey Corporation, )
) )

Defendants. )
)

)

PUBLIC VERSION
Exhibit 36
to the Declaration of Cullen N. Pendleton in Support of Amgen's Opposition to Roche's Motion for Summary Judgment that Claim 7 of the '349 Patent is Invalid Under 35 § USC 112 and is Not Infringed

Dockets.Justia.com

Case 1:05-cv-12237-WGY

Document 728-2

Filed 07/16/2007

Page 2 of 2

423

CONTROL OF MATERIALS

IRedacted I

IDENTITY OF THE WORKING CELL BANK (WCB) 29.04.93: PHENOTYPIC ANALYSIS

1. SUMMARY
Phenotypic propertes of the Working Cell Bank (WCB) 29.04.93 derived Chinese
hamster ovary (CHO) cells were analyzed by testing methotrexate (MTX)

resistance. The presence of the dihydrofolate reductase (DHFR) gene, correlated to the MTX resistance, was shown in the end-of-production cells (EPC) by their abilty to grow in the presence of MTX.

For this purpose CHO cells were harvested I I as EPC from a 10 L-Iaboratory scale fermentation. During this 10 L-Iaboratory scale
fermentation the CHO cells were cultivated in the absence of MTX which reflects the full scale fermentation process. The EPC were grown I I
I i in spinner flasks without MTX. Then the EPC were split into
cultivated for additional I I cycles to evaluate MTX resistance.

cultures without and with i I MTX in the medium. The EPC were then
Without MTX the specific Epoetin beta (EPO) productivity amounted to about

3.4 ¡.g EPOI1 06 cells/day I I. During furter cultivation without
productivity. Thus, in the extended cultivation

MTX the specific productivity decreased to about 50 % of the initial specific EPO
specific productivity of about 2 ¡.g EPO/1 06 cells! day was found. However in the presence of MTX the specifc productvity remained at a constant level of about

ILa

3.7¡.g EP0l106 cells/day.
In respect to growth expressed as doubling times the WCB 29.04.93 derived

CHO cells showed the same behavior as the CHO cells used for the serum
containing EPO production process.

Regarding specific productivity the WCB 29.04.93 derived CHO cells showed in

the absence of MTX the same reduction I I of the original specific
EPO productivity as the CHO cells used for the serum containing EPO

production process.

2. OBJECTIVE
The medium for the cultivation of the CHO cells contains no nucleosides. For the
biosynthesis of nucleosides the CHO cells have to synthesize appropriate

amounts of the enzyme dihydrofolate reductase (DHFR) for cell growt. MTX inhibits the DHFR. Thus, only if the gene coding for DHFR is available in
adequate copy numbers the cells can grow in the presence of MTX.

The objective of the phenotypic analysis was to prove MTX resistance of
WCB 29.04.93 derived CHO cells in extended serum-free culture.
Referenc: cm06645 Verion: 1.0
b-Ro.053821_3.2.S.2.3.MA T.wcb.mt

'Pharma Biotechnolog

1/5

Haussm~Dn Anja