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5 W* F. R. X) p' U6 f METHODS IN MOLECULAR BIOLOGY
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Protein Expression in Mammalian Cells
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5 J. |' j5 n% O8 a" s1 TMethods and Protocols
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# O/ y1 J4 c+ _Edited by& A/ M: B A% r- N' h4 |. F
James L. Hartley
0 g7 C1 j x" H: M. f9 aProtein Expression Laboratory, SAIC-Frederick, Inc., National Cancer Institute, * L8 o8 P6 y4 L) u: T2 i
Frederick, MD, USA
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* ^2 p, k$ H- A; LSpringer Science+Business Media, LLC 2012
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% L4 c- F N. m9 i. j2 P+ L" w- O$ RContents0 n) O# O/ F3 n' I: h2 u
Contents
* r5 s7 l2 I: m8 d( XPreface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
3 B# ?7 X: B! `7 ?Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix% X2 `8 g$ Q. d& I$ A( [
1 Why Proteins in Mammalian Cells? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
2 ]# f Q; r2 I9 AJames L. Hartley
, o+ k6 ` o. e1 x 2 Large-Scale Transfection of Mammalian Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13; Q) K3 W1 o3 y3 G" T p6 x
Lucia Baldi, David L. Hacker, Carine Meerschman,
3 {8 u ]: d( c# yand Florian M. Wurm
. |, G4 d- X+ y: l" L' v 3 Selection of High Expressing Mammalian Cells by Surface Display - X5 j" C) u4 p( U7 W' @
of Reporters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27, U( e r% \# T/ l' z# f! r$ A
Christine T. DeMaria
5 S. ^- k: i; |" A) b. i 4 Expression of a Secreted Protein in Mammalian Cells Using + [$ s4 L6 k. e' s: `4 y
Baculovirus Particles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
b( R7 d- ]& ^- ~5 kBarbara Ann Jardin, Cynthia B. Elias, and Satya Prakash
# M6 |8 P/ @2 S1 l2 R0 y5 m 5 Transfection of Difficult-to-Transfect Primary Mammalian Cells . . . . . . . . . . . . . . . 65
' l- U4 R, `3 L( }% d$ P9 NOliver Gresch and Ludger Altrogge
1 {5 ?3 G. p6 A2 n, D- q2 F 6 Stable Protein Expression in Mammalian Cells Using Baculoviruses . . . . . . . . . . . . 75
4 C" r. s7 O; R5 c+ n/ tAndreas Lackner, Emanuel Kreidl, Barbara Peter-Vörösmarty, & r" E( k( u+ y) D# T1 p
Sabine Spiegl-Kreinecker, Walter Berger, and Michael Grusch9 V( n7 i3 f5 F3 [' |* u, o6 S
7 Using Matrix Attachment Regions to Improve Recombinant
. H; r" N* \3 uProtein Production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
: ?% d1 p7 l# ?. ?* ]Niamh Harraghy, Montserrat Buceta, Alexandre Regamey, 0 t- |4 L2 ?7 Q* ]2 c
Pierre-Alain Girod, and Nicolas Mermod8 @1 K1 s7 _& n; t6 B6 Z6 ?# @
8 Controlling Apoptosis to Optimize Yields of Proteins from Mammalian Cells . . . . . 111
k' a0 u# o4 G' I; R, [: dMatthew P. Zustiak, Haimanti Dorai, Michael J. Betenbaugh,
- T. P- a8 U+ T7 z% f! F' J- Pand Tina M. Sauerwald
) n. b+ f( D( ?" N 9 Post-transcriptional Regulatory Elements for Enhancing Transient Gene
" x, `, P8 p- C ~3 f! _Expression Levels in Mammalian Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1251 L* A0 w; U7 T" g% }
Mariati, Steven C.L. Ho, Miranda G.S. Yap, and Yuansheng Yang% D$ c6 ]8 `1 R ]
10 Converting Monoclonal Antibodies into Fab Fragments for Transient
- R( K. d+ l' C( U B x; XExpression in Mammalian Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
% w3 a6 m8 Q8 B) ~9 l: K! VJoanne E. Nettleship, Aleksandra Flanagan, Nahid Rahman-Huq, : Y& V( M- n- P
Rebecca Hamer, and Raymond J. Owens
. e+ p; h6 D) t9 v; [" Z11 Generation of High-Expressing Cells by Methotrexate Amplification
# e: h( Y8 E5 Iof Destabilized Dihydrofolate Reductase Selection Marker . . . . . . . . . . . . . . . . . . . 161# l( b- \; h" m; Z
Say Kong Ng
3 T- O" n( z5 u. [12 Tools for Coproducing Multiple Proteins in Mammalian Cells . . . . . . . . . . . . . . . . 173
3 j& K9 @( L( M0 U9 X# D2 o. VZahra Assur, Wayne A. Hendrickson, and Filippo Mancia
/ J6 \; @: P* V# J2 Z# m/ x5 ?! c5 i0 jviii Contents
; O9 ~4 j/ T6 l13 Identification and Characterization of Protein Glycosylation Using Specific 4 F" h q c: f8 X4 l% l
Endo- and Exoglycosidases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
+ \# \. y+ ]+ i6 PPaula Magnelli, Alicia Bielik, and Ellen Guthrie' z, D) H+ ?3 F7 K$ h
14 Strategies for Efficient Transfection of CHO-Cells with Plasmid DNA . . . . . . . . . . 2139 \7 L7 A2 k S) n; S; s, d
Renate Kunert and Karola Vorauer-Uhl
4 j/ f9 P8 Q+ C* i15 Methods for Constructing Clones for Protein Expression in Mammalian Cells . . . . 227
" L* _% f& H' B& O) N5 {2 C! FTakefumi Sone and Fumio Imamoto
- L4 Z) `! x, a' f( e16 Optimizing Transient Recombinant Protein Expression in Mammalian Cells . . . . . . 251; O2 t( ~- H9 o" b4 [
Ralph F. Hopkins, Vanessa E. Wall, and Dominic Esposito0 U- o4 E6 r+ M' U
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 269
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4 |. F/ L2 @; r% C" C* F8 V[hide][/hide]
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发表于 2011-12-5 09:21
