A chimeric anti-CEA antibody with heavy interchain disulfide bonds deleted: Molecular characterization and biodistributions in normal and tumor bearing mice
M. Neumaier et al., A chimeric anti-CEA antibody with heavy interchain disulfide bonds deleted: Molecular characterization and biodistributions in normal and tumor bearing mice, ANTICANC R, 19(1A), 1999, pp. 13-21
We have deleted the interchain disulfide bonds in a chimeric anti-CEA antib
ody (chT84.66) by mutating two cysteines in the heavy chain to glycine resi
dues. The resulting antibody Delta SSchT84.66 was expressed in high yield i
n a bioreactor and purified to homogeneity in a single step on an anti-idio
typic antibody affinity column. The molecular size of the antibody was 150
kDa as judged by gel filtration, SDS gel electrophoresis under non-reducing
conditions, and MALDITOF/MS. The 150 kDa antibody had nearly identical kin
etic (k(on) = 1.53 x 10(6) M-1 s(-1),.k(off) = 1.14 x 10(-5) s(-1)) and aff
inity constants (K-aff = 1.34 x 10 M-1) compared to the parent murine (K-af
f = 1.25 x 10(11) M-1) and chimeric (K-aff =1.16 x 10(11) M-1) antibodies w
hen tested on biosensor chips. When Delta SSchT84.66 was conjugated to the
isothiocynato derivative of DTPA, radiolabeled with In-111, and injected in
to either noimnl ol nude mice bearing tumor xenografts, it gave nearly iden
tical biodistributions to chT84.66. Delta SSchT84.66 and chT84.66 antibodie
s gave a maximum tumor uptake of 48 and 74 %ID/g, and tumor to blood ratios
of 5.3 and 6.2 at 48 h, respectively. We conclude that Delta SSchT84.66 ir
reversibly associates into H2L2 dimers after concentration, that the dimers
are stable under both the in vitro and in vivo conditions used in this stu
dy, and the properties of the antibody are virtually indistinguishable from
the parent chT84.66 antibody.