Heterodimeric antibody fc-containing proteins and methods for

110 Pages·2013·5.68 MB·English

US 20130039913A1
(19) United States
(12) Patent Application Publication (10) Pub. No.: US 2013/0039913 A1
Labrijn et al. (43) Pub. Date: Feb. 14, 2013
(54) HETERODIMERIC ANTIBODY (30) Foreign Application Priority Data
FC-CONTAINING PROTEINS AND METHODS
FOR PRODUCTION THEREOF Apr. 20, 2010 (DK) ........................... .. PA201000330
(75) Inventors: Aran Frank Labrijn, Nigtevecht (NL); Nov. 24, 2010 (DK) ........................... .. PA201001066
Joyce I. Meesters, Utrecht (NL); EWald
T. J. Van Den Bremer, Best (NL); Joost
J. Neijssen, Werkhoven (NL); Patrick Publication Classi?cation
Van Berkel, Utrecht (NL); Bart De
Goeij, Maarssen (NL); Tom Vink, (51) Int. Cl.
Alphen aan den Rijn (NL); Jan Van De A61K 39/395 (2006.01)
Winkel, Zeist (NL); Janine C12N 5/10 (2006.01)
Schuurman, Diemen (NL); Paul A61P 35/00 (2006.01)
Parren, Odijk (NL) G01N 33/566 (2006.01)
(73) Assignee: GENMAB A/S, Copenhagen (DK) C07K 16/18 (2006.01)
C12P 21/00 (2006.01)
(21) Appl. No.: 13/642,253 C12N 15/63 (2006.01)
(52) U.S. Cl. .................. .. 424/136.1; 435/69.6; 435/352;
(22) PCT Filed: Apr. 20, 2011 435/358; 435/320.1; 436/501; 530/387.3;
530/391.7
(86) PCT No.: PCT/EP11/56388
§ 371 (00)’
(57) ABSTRACT
(2), (4) Date: Oct. 24, 2012
Related U.S. Application Data
Heterodimeric antibody-Fc-containing proteins, such as
(60) Provisional application No. 61/326,082, ?led on Apr. bispeci?c antibodies, and novel methods for producing such
20, 2010. proteins.
Patent Application Publication Feb. 14, 2013 Sheet 1 0f 64 US 2013/0039913 A1
Figure 1
5
+ RhlgG4-2F8 X HuIgG4-7D8
E 4 <> HulgG4-2F8 x RhlgG4-7D8
g 3 + HulgG4-2F8 x HulgG4-7D8
g; + RhlgG4-2F8 X RhlgG4-7D8
Q 2
O
1
O
0.1 1 10 100 1000 10000
Concentration (ng/mL)
Patent Application Publication Feb. 14, 2013 Sheet 2 0f 64 US 2013/0039913 A1
Figure 2
Species Isotype Amino-acid position‘
(common name) Core-hinge CH$CH3 interface
Homosapiens IgG1 CPPCP QYTLTLKKTPVLDFYK
I962 - - - - - - - - - - - - - - - M - - - - -
I963 - - R - - - - - - - - - N - - M - - - - -
I964 - — S - - - - - - - - - - - - - - - - - R
Macaca mulatta lgG‘I » - - - - ~ - - - - - - - - - - - ~ - - -
(Rhesus Monkey) lgGZ ' - - - - - - - - - - - - - - - - - - - -
I963 - - - - - - - I - - - T - - - - - - - -
IIggGG44((ICnh))"‘ “ - - A - - - - l v w - T - - - - - - L - -
‘EU numbering; “In, Indian; "'Ch, Chinese
Patent Application Publication Feb. 14, 2013 Sheet 3 0f 64 US 2013/0039913 A1
'Fme (hours)
Patent Application Publication Feb. 14, 2013 Sheet 4 0f 64 US 2013/0039913 A1
‘lime (hours)
Patent Application Publication Feb. 14, 2013 Sheet 5 0f 64 US 2013/0039913 A1
Figure 5
5
—o- lgG4-2F8 x lgG4—7D8
4
E -I- lgG1-2F8-ITL x lgG4-7D8-CPPC
C
Lo 3
9r
2
D
O o
1
0
0:1 5 1'0 160
Concentration 2-MEA (mM)
Patent Application Publication Feb. 14, 2013 Sheet 6 0f 64 US 2013/0039913 A1
Figure 6
A
IgG1-2F8-ITL X IgG4-7D8-CPPC
1462599
145543.? 0 mM 2-MEA
145000 ' 146000 147000 148000 Da
145902.?
1452616
7 mM 2-MEA
1455448
1!
RIM} kilo-n
145000 146000 147000 148000Da
1459010
40 mM 2-MEA
1452503
145000 146000 147000 148000 Da
Patent Application Publication Feb. 14, 2013 Sheet 7 0f 64 US 2013/0039913 A1
Figure 6 (continued)
B%ispecifi c
20
0.1 1 1'0 150
Concentration 2-MEA (mM)
Patent Application Publication Feb. 14, 2013 Sheet 8 0f 64 US 2013/0039913 A1
Figure 7
A
lgG1-2F8-ITL x lgG4-7D8-CPPC
100'
Bb(ocioisnnptfder%cioinlf gc)